DOCSLIB.ORG

Mouse Acy1 Knockout Project (CRISPR/Cas9)

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mouse Acy1 Knockout Project (CRISPR/Cas9) https://www.alphaknockout.com Mouse Acy1 Knockout Project (CRISPR/Cas9) Objective: To create a Acy1 knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering. Strategy summary: The Acy1 gene (NCBI Reference Sequence: NM_025371 ; Ensembl: ENSMUSG00000023262 ) is located on Mouse chromosome 9. 15 exons are identified, with the ATG start codon in exon 2 and the TGA stop codon in exon 15 (Transcript: ENSMUST00000024031). Exon 2~15 will be selected as target site. Cas9 and gRNA will be co-injected into fertilized eggs for KO Mouse production. The pups will be genotyped by PCR followed by sequencing analysis. Note: Exon 2 starts from about 0.08% of the coding region. Exon 2~15 covers 100.0% of the coding region. The size of effective KO region: ~4637 bp. The KO region does not have any other known gene. Page 1 of 9 https://www.alphaknockout.com Overview of the Targeting Strategy Wildtype allele 5' gRNA region gRNA region 3' 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Legends Exon of mouse Acy1 Knockout region Page 2 of 9 https://www.alphaknockout.com Overview of the Dot Plot (up) Window size: 15 bp Forward Reverse Complement Sequence 12 Note: The 2000 bp section upstream of start codon is aligned with itself to determine if there are tandem repeats. Tandem repeats are found in the dot plot matrix. The gRNA site is selected outside of these tandem repeats. Overview of the Dot Plot (down) Window size: 15 bp Forward Reverse Complement Sequence 12 Note: The 2000 bp section downstream of stop codon is aligned with itself to determine if there are tandem repeats. No significant tandem repeat is found in the dot plot matrix. So this region is suitable for PCR screening or sequencing analysis. Page 3 of 9 https://www.alphaknockout.com Overview of the GC Content Distribution (up) Window size: 300 bp Sequence 12 Summary: Full Length(2000bp) | A(26.45% 529) | C(25.1% 502) | T(21.05% 421) | G(27.4% 548) Note: The 2000 bp section upstream of start codon is analyzed to determine the GC content. Significant high GC-content regions are found. The gRNA site is selected outside of these high GC-content regions. Overview of the GC Content Distribution (down) Window size: 300 bp Sequence 12 Summary: Full Length(2000bp) | A(24.0% 480) | C(28.85% 577) | T(24.2% 484) | G(22.95% 459) Note: The 2000 bp section downstream of stop codon is analyzed to determine the GC content. No significant high GC-content region is found. So this region is suitable for PCR screening or sequencing analysis. Page 4 of 9 https://www.alphaknockout.com BLAT Search Results (up) QUERY SCORE START END QSIZE IDENTITY CHROM STRAND START END SPAN ----------------------------------------------------------------------------------------------- browser details YourSeq 2000 1 2000 2000 100.0% chr9 - 106437723 106439722 2000 browser details YourSeq 325 48 408 2000 95.3% chr17 - 45655599 45655943 345 browser details YourSeq 306 36 409 2000 92.9% chr12 + 62931214 62931585 372 browser details YourSeq 270 36 408 2000 90.7% chr2 - 30145534 30146003 470 browser details YourSeq 269 35 404 2000 88.5% chr7 - 127694524 127694858 335 browser details YourSeq 268 36 409 2000 87.9% chr10 - 78431255 78431598 344 browser details YourSeq 268 39 408 2000 92.6% chr1 + 39320733 39321128 396 browser details YourSeq 264 45 404 2000 90.4% chr18 + 20799349 20991849 192501 browser details YourSeq 261 46 398 2000 88.9% chr12 - 69604724 69605042 319 browser details YourSeq 259 36 398 2000 88.6% chr19 + 7313169 7313493 325 browser details YourSeq 256 45 412 2000 88.1% chr6 - 129360634 129360975 342 browser details YourSeq 254 36 404 2000 86.9% chr2 - 128725446 128725786 341 browser details YourSeq 254 36 397 2000 87.1% chr11 - 95933618 95933947 330 browser details YourSeq 251 45 398 2000 91.0% chr12 + 54149701 54583824 434124 browser details YourSeq 250 35 404 2000 87.2% chr14 - 63866608 63866926 319 browser details YourSeq 250 36 392 2000 89.1% chr19 + 7006212 7006538 327 browser details YourSeq 249 36 408 2000 86.2% chr17 - 12807411 12807754 344 browser details YourSeq 249 36 397 2000 88.7% chr12 + 112418932 112419260 329 browser details YourSeq 248 35 398 2000 86.3% chrX - 7225694 7226024 331 browser details YourSeq 248 60 404 2000 88.3% chr7 + 46494666 46494982 317 Note: The 2000 bp section upstream of start codon is BLAT searched against the genome. No significant similarity is found. BLAT Search Results (down) QUERY SCORE START END QSIZE IDENTITY CHROM STRAND START END SPAN ----------------------------------------------------------------------------------------------- browser details YourSeq 2000 1 2000 2000 100.0% chr9 - 106431084 106433083 2000 browser details YourSeq 477 1516 2000 2000 99.2% chr5 + 115801558 115802042 485 browser details YourSeq 476 1517 2000 2000 99.2% chr9 - 7751842 7752325 484 browser details YourSeq 475 1516 2000 2000 99.0% chr2 - 134804063 134804547 485 browser details YourSeq 475 1516 2000 2000 99.0% chr1 - 43189273 43189757 485 browser details YourSeq 472 1521 2000 2000 99.2% chr13 - 55085894 55086373 480 browser details YourSeq 471 1516 2000 2000 98.6% chr1 - 134518198 134518682 485 browser details YourSeq 471 1516 2000 2000 98.6% chr1 + 179520256 179520740 485 browser details YourSeq 469 1517 2000 2000 98.6% chr12 - 11099711 11100195 485 browser details YourSeq 465 1516 2000 2000 97.6% chr2 - 102174393 102174876 484 browser details YourSeq 459 1518 2000 2000 97.6% chr18 - 34737532 34738014 483 browser details YourSeq 457 1522 2000 2000 97.8% chr4 - 55254854 55255332 479 browser details YourSeq 456 1516 2000 2000 97.2% chr6 + 54177079 54177566 488 browser details YourSeq 455 1516 2000 2000 97.0% chr13 - 4614297 4614781 485 browser details YourSeq 452 1516 2000 2000 95.9% chr10 - 116631943 116632422 480 browser details YourSeq 451 1517 2000 2000 96.0% chr3 + 53401610 53402083 474 browser details YourSeq 450 1517 2000 2000 95.9% chr18 + 30458965 30459445 481 browser details YourSeq 445 1521 2000 2000 96.2% chr18 - 67562504 67562965 462 browser details YourSeq 442 1516 1997 2000 95.9% chr12 + 103027825 103028292 468 browser details YourSeq 441 1518 2000 2000 95.4% chr15 - 96188744 96189222 479 Note: The 2000 bp section downstream of stop codon is BLAT searched against the genome. No significant similarity is found. Page 5 of 9 https://www.alphaknockout.com Gene and protein information: Acy1 aminoacylase 1 [ Mus musculus (house mouse) ] Gene ID: 109652, updated on 12-Aug-2019 Gene summary Official Symbol Acy1 provided by MGI Official Full Name aminoacylase 1 provided by MGI Primary source MGI:MGI:87913 See related Ensembl:ENSMUSG00000023262 Gene type protein coding RefSeq status VALIDATED Organism Mus musculus Lineage Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus Also known as Acy-1; 1110014J22Rik Expression Broad expression in duodenum adult (RPKM 66.0), kidney adult (RPKM 51.7) and 19 other tissues See more Orthologs human all Genomic context Location: 9 F1; 9 57.49 cM See Acy1 in Genome Data Viewer Exon count: 15 Annotation release Status Assembly Chr Location 108 current GRCm38.p6 (GCF_000001635.26) 9 NC_000075.6 (106432981..106438236, complement) Build 37.2 previous assembly MGSCv37 (GCF_000001635.18) 9 NC_000075.5 (106335327..106340567, complement) Chromosome 9 - NC_000075.6 Page 6 of 9 https://www.alphaknockout.com Transcript information: This gene has 12 transcripts Gene: Acy1 ENSMUSG00000023262 Description aminoacylase 1 [Source:MGI Symbol;Acc:MGI:87913] Gene Synonyms 1110014J22Rik, Acy-1 Location Chromosome 9: 106,432,981-106,438,319 reverse strand. GRCm38:CM001002.2 About this gene This gene has 12 transcripts (splice variants), 209 orthologues, 3 paralogues and is a member of 1 Ensembl protein family. Transcripts Name Transcript ID bp Protein Translation ID Biotype CCDS UniProt Flags Acy1- ENSMUST00000024031.12 1529 408aa ENSMUSP00000024031.6 Protein coding CCDS23477 A0A0R4J050 TSL:1 201 GENCODE basic APPRIS P1 Acy1- ENSMUST00000214275.1 1284 373aa ENSMUSP00000149789.1 Protein coding - A0A1L1SS90 TSL:5 208 GENCODE basic Acy1- ENSMUST00000215395.1 1051 343aa ENSMUSP00000149394.1 Protein coding - A0A1L1SRC1 TSL:5 209 GENCODE basic Acy1- ENSMUST00000216400.1 1030 336aa ENSMUSP00000149636.1 Protein coding - A0A1L1SRW7 TSL:5 211 GENCODE basic Acy1- ENSMUST00000190972.2 855 219aa ENSMUSP00000139953.1 Protein coding - A0A087WPX1 CDS 3' 207 incomplete TSL:3 Acy1- ENSMUST00000190900.1 616 50aa ENSMUSP00000140582.1 Protein coding - A0A087WRE0 CDS 3' 206 incomplete TSL:3 Acy1- ENSMUST00000217531.1 466 155aa ENSMUSP00000149516.1 Protein coding - A0A1L1SRL5 CDS 5' and 3' 212 incomplete TSL:5 Acy1- ENSMUST00000215506.1 1122 201aa ENSMUSP00000150039.1 Nonsense mediated - A0A1L1SSU4 TSL:5 210 decay Acy1- ENSMUST00000190803.1 771 39aa ENSMUSP00000139931.1 Nonsense mediated - A0A087WPV3 TSL:5 204 decay Acy1- ENSMUST00000189097.6 1885 No - Retained intron - - TSL:5 203 protein Acy1- ENSMUST00000187324.7 1832 No - Retained intron - - TSL:5 202 protein Acy1- ENSMUST00000190851.2 1467 No - Retained intron - - TSL:5 205 protein Page 7 of 9 https://www.alphaknockout.com 25.34 kb Forward strand 106.425Mb 106.430Mb 106.435Mb 106.440Mb 106.445Mb Genes Rpl29-203 >protein coding Abhd14b-209 >protein coding (Comprehensive set... Rpl29-202 >protein coding Abhd14b-208 >protein coding Rpl29-201 >protein coding Abhd14b-205 >protein coding Rpl29-204 >protein coding Abhd14b-206 >protein coding Rpl29-206 >protein coding Abhd14b-204 >protein coding Rpl29-205 >protein coding
Load more

Recommended publications
  • Human ACY1 / Aminoacylase-1 Protein (His Tag)
    Human ACY1 / Aminoacylase-1 Protein (His Tag) Catalog Number: 10549-H08B General Information SDS-PAGE: Gene Name Synonym: ACY-1; ACY1D; HEL-S-5 Protein Construction: A DNA sequence encoding the full length of human ACY1 (NP_000657.1) (Met 1-Ser 408) was expressed with a polyhistidine tag at the C-terminus. Source: Human Expression Host: Baculovirus-Insect Cells QC Testing Purity: > 95 % as determined by SDS-PAGE Endotoxin: Protein Description < 1.0 EU per μg of the protein as determined by the LAL method Aminoacylase 1 (ACY1), a metalloenzyme that removes amide-linked Stability: ACY1 groups from amino acids and may play a role in regulating responses to oxidative stress. Both the C-terminal fragment found in the Samples are stable for up to twelve months from date of receipt at -70 ℃ two-hybrid screen and full-length ACY1 co-immunoprecipitate with SphK1. Though both C-terminal and full-length proteins slightly reduce SphK1 Predicted N terminal: Met activity measured in vitro, the C-terminal fragment inhibits while full-length Molecular Mass: ACY1 potentiates the effects of SphK1 on proliferation and apoptosis. It suggested that ACY1 physically interacts with SphK1 and may influence its The recombinant human ACY1 consists of 419 amino acids and predicts a physiological functions. As a homodimeric zinc-binding enzyme, molecular mass of 47.3 kDa. It migrates as an approximately 44 kDa Aminoacylase 1 catalyzes the hydrolysis of N alpha-acylated amino acids. protein in SDS-PAGE under reducing conditions. Deficiency of Aminoacylase 1 due to mutations in the Aminoacylase 1 (ACY1) gene follows an autosomal-recessive trait of inheritance and is Formulation: characterized by accumulation of N-acetyl amino acids in the urine.
    [Show full text]
  • Analysis of the Complex Interaction of Cdr1as‑Mirna‑Protein and Detection of Its Novel Role in Melanoma
    ONCOLOGY LETTERS 16: 1219-1225, 2018 Analysis of the complex interaction of CDR1as‑miRNA‑protein and detection of its novel role in melanoma LIHUAN ZHANG*, YUAN LI*, WENYAN LIU, HUIFENG LI and ZHIWEI ZHU College of Life Sciences, Shanxi Agricultural University, Taigu, Shanxi 030801, P.R. China Received May 15, 2017; Accepted April 9, 2018 DOI: 10.3892/ol.2018.8700 Abstract. Despite improvements in the prevention, diagnosis has been detected in several species, including viruses (2), and treatment of melanoma having developed rapidly, the role plants (4), archaea (5)[Salzman, 2013 #198; Memczak, 2013 of circular RNA CDR1 antisense RNA (CDR1as) in melanoma #291] and animals (6). In eukaryotic cells, circRNA has remains to be elucidated. The aim of the present study was to attracted attention due to its unique characteristics, including predict the novel roles of CDR1as in melanoma through novel high stability, specificity and evolutionary conservation (7). bioinformatics analysis. In the present study, the circ2Traits The majority of circRNAs are derived from exons of coding database was used to supply information on CDR1as in cancer. regions, 3'UTRs, 5'UTRs, introns, intergenetic regions and CircNet, circBase and circInteractome databases were used to antisense RNAs (8). CircRNAs, which have attracted attention detect the co-expression of CDR1as, microRNAs and proteins. in recent years, can be produced by canonical and nonca- Furthermore, the functions and pathways of the associated nonical splicing as distinguished from the linear RNAs (9). proteins were predicted using the Database for Annotation, Through high-throughput sequencing, three types of circRNAs Visualization and Integrated Discovery. Gene Ontology have been identified: Exonic circRNAs (7), circular intronic enrichment analysis suggested that the proteins associated RNAs (ciRNAs) (9), and retained-intron circular RNAs or with CDR1as were mainly regulated in the cytoplasm as exon-intron circRNAs (elciRNAs) (10).
    [Show full text]
  • Polymorphic Human Somatostatin Gene Is Located on Chromosome 3 (Gene Mapping/Somatic Cell Hybrids/DNA Polymorphism) SUSAN L
    Proc. NatL Acad. Sci. USA Vol. 80, pp. 2686-2689, May 1983 Genetics Polymorphic human somatostatin gene is located on chromosome 3 (gene mapping/somatic cell hybrids/DNA polymorphism) SUSAN L. NAYLOR*, ALAN Y. SAKAGUCHI*, LU-PING SHENtt, GRAEME I. BELLO, WILLIAM J. RUTTERt, AND THOMAS B. SHOWS* *Department of Human Genetics, Roswell Park Memorial Institute, New York State Department of Health, Buffalo, New York 14263; and tDepartment of Biochemistry and Biophysics, University of California, San Francisco, California 94143 Communicated by Harry Harris, January 20, 1983 ABSTRACT Somatostatin is a 14-amino-acid neuropeptide and mosomes are given in the legend to Table 2. Most relevant to hormone that inhibits the secretion of several peptide hormones. this study are two series of hybrids having translocations with The human gene for somatostatin SST has been cloned, and the chromosome 3. TSL hybrids, resulting from the fusion of the sequence has been determined. This clone was used as a probe human fibroblast line GM2808 [46,XX,t(3;17)(p21;p13)] with in chromosome mapping studies to detect the human somatostatin mouse LMTK- cells, segregate 3/17 translocation chromo- sequence in human-rodent hybrids. Southern blot analysis of 41 somes. Only those hybrids retaining the 17/3 translocation hybrids, including some containing translocations of human chro- chromosome (17qterl7pl3::3p21-+3qter) or a normal chro- mosomes, placed SST in the q21--qter region of chromosome 3. mosome 17 having the thymidine kinase gene proliferated on Human DNAs from unrelated individuals were screened for re- hypoxanthine/aminopterin/thymidine selection medium (10). striction fragment polymorphisms detectable by the somatostatin XTR was made from a fusion gene probe.
    [Show full text]
  • Human ACY1 ELISA Matched Antibody Pair
    Version 01-06/20 User's Manual Human ACY1 ELISA Matched Antibody Pair ABPR-0012 This product is for research use only and is not intended for diagnostic use. For illustrative purposes only. To perform the assay the instructions for use provided with the kit have to be used. Creative Diagnostics Address: 45-1 Ramsey Road, Shirley, NY 11967, USA Tel: 1-631-624-4882 (USA) 44-161-818-6441 (Europe) Fax: 1-631-938-8221 Email: [email protected] Web: www.creative-diagnostics.com Cat: ABPR-0012 Human ACY1 ELISA Matched Antibody Pair Version 21-06/20 PRODUCT INFORMATION Intended Use This antibody pair set comes with matched antibody pair to detect and quantify protein level of human ACY1. General Description This gene encodes a cytosolic, homodimeric, zinc-binding enzyme that catalyzes the hydrolysis of acylated L- amino acids to L-amino acids and an acyl group, and has been postulated to function in the catabolism and salvage of acylated amino acids. This gene is located on chromosome 3p21.1, a region reduced to homozygosity in small-cell lung cancer (SCLC), and its expression has been reported to be reduced or undetectable in SCLC cell lines and tumors. The amino acid sequence of human aminoacylase-1 is highly homologous to the porcine counterpart, and this enzyme is the first member of a new family of zinc-binding enzymes. Mutations in this gene cause aminoacylase-1 deficiency, a metabolic disorder characterized by central nervous system defects and increased urinary excretion of N-acetylated amino acids. Alternative splicing of this gene results in multiple transcript variants.
    [Show full text]
  • Functional Annotation of the Human Retinal Pigment Epithelium
    BMC Genomics BioMed Central Research article Open Access Functional annotation of the human retinal pigment epithelium transcriptome Judith C Booij1, Simone van Soest1, Sigrid MA Swagemakers2,3, Anke HW Essing1, Annemieke JMH Verkerk2, Peter J van der Spek2, Theo GMF Gorgels1 and Arthur AB Bergen*1,4 Address: 1Department of Molecular Ophthalmogenetics, Netherlands Institute for Neuroscience (NIN), an institute of the Royal Netherlands Academy of Arts and Sciences (KNAW), Meibergdreef 47, 1105 BA Amsterdam, the Netherlands (NL), 2Department of Bioinformatics, Erasmus Medical Center, 3015 GE Rotterdam, the Netherlands, 3Department of Genetics, Erasmus Medical Center, 3015 GE Rotterdam, the Netherlands and 4Department of Clinical Genetics, Academic Medical Centre Amsterdam, the Netherlands Email: Judith C Booij - [email protected]; Simone van Soest - [email protected]; Sigrid MA Swagemakers - [email protected]; Anke HW Essing - [email protected]; Annemieke JMH Verkerk - [email protected]; Peter J van der Spek - [email protected]; Theo GMF Gorgels - [email protected]; Arthur AB Bergen* - [email protected] * Corresponding author Published: 20 April 2009 Received: 10 July 2008 Accepted: 20 April 2009 BMC Genomics 2009, 10:164 doi:10.1186/1471-2164-10-164 This article is available from: http://www.biomedcentral.com/1471-2164/10/164 © 2009 Booij et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
    [Show full text]
  • Study of the Expression and Function of ACY1 in Patients with Colorectal Cancer
    ONCOLOGY LETTERS 13: 2459-2464, 2017 Study of the expression and function of ACY1 in patients with colorectal cancer BING YU1,2, XUEZHONG LIU3, XIUZHEN CAO2, MINGYUE ZHANG2 and HONG CHANG1 1Department of Hepatobiliary Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021; 2Department of Colorectal Surgery, Tai'an Central Hospital, Tai'an, Shandong 271000; 3Department of Gastrointestinal Surgery, Liaocheng People's Hospital, Liaocheng, Shandong 252000, P.R. China Received February 22, 2016; Accepted October 19, 2016 DOI: 10.3892/ol.2017.5702 Abstract. Aminoacylase 1 (ACY1) is important for regulating area of intense study; however, it is a complex issue. Previous the proliferation of numerous types of cancer. However, the studies indicate that tumor cells of different clinical stages expression and mechanisms underlying the function of ACY1 may exhibit different expression levels of certain biomarkers, in colorectal cancer remain unclear. In order to investigate which could be used as a target for the diagnosis or treatment the expression and function of ACY1 in colorectal cancer, of the tumor (3-5). In addition, with ongoing and advancing tumor tissue and blood samples were collected for analysis research into tumor biology, new targets are being identified from 132 patients diagnosed with colorectal cancer. Reverse and studied, including aminoacylase 1 (ACY1) (6). transcription-quantitative polymerase chain reaction analysis ACY1 is a cytosolic enzyme that deacylates the α-acylated and western blotting identified significantly increased expres- amino acid from the N-terminal peptide of intracellular sion of ACY1 mRNA in colorectal tumor tissue (P<0.05 vs. proteins (6). Following this, terminal acetylated proteins are adjacent normal tissue) and notably increased ACY1 protein more stable (7-9).
    [Show full text]
  • Human Aminoacylase/ACY1 Antibody Catalog Number: ATGA0329
    Human Aminoacylase/ACY1 antibody Catalog Number: ATGA0329 PRODUCT INPORMATION Catalog number ATGA0329 Clone No. AT1E2 Product type Monoclonal Antibody UnitProt No. Q03154 NCBI Accession No. NP_000657 Alternative Names Aminoacylase 1, ACY1D, ACYLASE, Aminoacylase 1 PRODUCT SPECIFICATION Antibody Host Mouse Reacts With Human Concentration 1mg/ml (determined by BCA assay) Formulation Liquid in. Phosphate-Buffered Saline (pH 7.4) with 0.02% Sodium Azide, 10% glycerol Immunogen Recombinant human ACY1 (1-408aa) purified from E. coli Isotype IgG2b kappa Purification Note By protein-A affinity chromatography Application ELISA,WB,ICC/IF,FACS Usage The antibody has been tested by ELISA, Western blot, ICC/IF and FACS analysis to assure specificity and reactivity. Since application varies, however, each investigation should be titrated by the reagent to obtain optimal results. 1 Human Aminoacylase/ACY1 antibody Catalog Number: ATGA0329 Storage Can be stored at +2C to +8C for 1 week. For long term storage, aliquot and store at -20C to -80C. Avoid repeated freezing and thawing cycles. BACKGROUND Description Aminoacylase-1, also designated N-acyl-L-amino-acid amidohydrolase or ACY1, is a member of the largest metallopeptidase family called M20A. Aminoacylase-1 is a zinc-binding homodimeric enzyme expressed in kidney, brain, placenta and spleen. It is the most abundant of the aminoacylases. Defects in ACY1 are the cause of aminoacylase-1 deficiency (ACY1D). ACY1D results in a metabolic disorder manifesting with encephalopathy, unspecific psychomotor delay, psychomotor delay with atrophy of the vermis and syringomyelia, marked muscular hypotonia or normal clinical features. All affected individuals exhibit markedly increased urinary excretion of several N-acetylated amino acids.
    [Show full text]
  • Anti-ACY1 / Aminoacylase 1 Antibody (ARG40257)
    Product datasheet [email protected] ARG40257 Package: 100 μl anti-ACY1 / Aminoacylase 1 antibody Store at: -20°C Summary Product Description Rabbit Polyclonal antibody recognizes ACY1 / Aminoacylase 1 Tested Reactivity Hu, Ms, Rat Tested Application ICC/IF, IHC-P, WB Host Rabbit Clonality Polyclonal Isotype IgG Target Name ACY1 / Aminoacylase 1 Antigen Species Human Immunogen Recombinant fusion protein corresponding to aa. 1-408 of Human ACY1 / Aminoacylase 1 (NP_001185824.1). Conjugation Un-conjugated Alternate Names ACY-1; N-acyl-L-amino-acid amidohydrolase; ACY1D; EC 3.5.1.14; HEL-S-5; Aminoacylase-1 Application Instructions Application table Application Dilution ICC/IF 1:50 - 1:200 IHC-P 1:50 - 1:200 WB 1:500 - 1:2000 Application Note * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. Positive Control A431, Mouse kidney and Rat kidney Calculated Mw 46 kDa Observed Size ~ 45 kDa Properties Form Liquid Purification Affinity purified. Buffer PBS (pH 7.3), 0.02% Sodium azide and 50% Glycerol. Preservative 0.02% Sodium azide Stabilizer 50% Glycerol Storage instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw www.arigobio.com 1/3 cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use. Note For laboratory research only, not for drug, diagnostic or other use. Bioinformation Gene Symbol ACY1 Gene Full Name aminoacylase 1 Background This gene encodes a cytosolic, homodimeric, zinc-binding enzyme that catalyzes the hydrolysis of acylated L-amino acids to L-amino acids and an acyl group, and has been postulated to function in the catabolism and salvage of acylated amino acids.
    [Show full text]
  • Human Aminoacylase/ACY1 Antibody Antigen Affinity-Purified Polyclonal Goat Igg Catalog Number: AF2900
    Human Aminoacylase/ACY1 Antibody Antigen Affinity-purified Polyclonal Goat IgG Catalog Number: AF2900 DESCRIPTION Species Reactivity Human Specificity Detects human and mouse Aminoacylase/ACY1 in direct ELISAs and Western blots. Source Polyclonal Goat IgG Purification Antigen Affinity­purified Immunogen S. frugiperda insect ovarian cell line Sf 21­derived recombinant human Aminoacylase/ACY1 Thr2­Ser408 Accession # Q03154 Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. *Small pack size (­SP) is supplied either lyophilized or as a 0.2 μm filtered solution in PBS. APPLICATIONS Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website. Recommended Sample Concentration Western Blot 0.1 µg/mL Recombinant Human Aminoacylase/ACY1 (Catalog # 2900­ZN) Immunoprecipitation 25 µg/mL Conditioned cell culture medium spiked with Recombinant Human Aminoacylase/ACY1 (Catalog # 2900­ZN), see our available Western blot detection antibodies PREPARATION AND STORAGE Reconstitution Reconstitute at 0.2 mg/mL in sterile PBS. Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (­SP) is shipped with polar packs. Upon receipt, store it immediately at ­20 to ­70 °C Stability & Storage Use a manual defrost freezer and avoid repeated freeze­thaw cycles. l 12 months from date of receipt, ­20 to ­70 °C as supplied. l 1 month, 2 to 8 °C under sterile conditions after reconstitution. l 6 months, ­20 to ­70 °C under sterile conditions after reconstitution. BACKGROUND The human ACY1 gene encodes Aminoacylase, a member of the M20 family of metalloproteases (1).
    [Show full text]
  • ACY1 Polyclonal Antibody Product Information
    ACY1 Polyclonal Antibody Cat #: ABP57702 Size: 30μl /100μl /200μl Product Information Product Name: ACY1 Polyclonal Antibody Applications: WB, ELISA Isotype: Rabbit IgG Reactivity: Human, Mouse Catalog Number: ABP57702 Lot Number: Refer to product label Formulation: Liquid Concentration: 1 mg/ml Storage: Store at -20°C. Avoid repeated Note: Contain sodium azide. freeze / thaw cycles. Background: ACY1 encodes a cytosolic, homodimeric, zinc-binding enzyme that catalyzes the hydrolysis of acylated L-amino acids to L-amino acids and an acyl group, and has been postulated to function in the catabolism and salvage of acylated amino acids. ACY1 is located on chromosome 3p21. , a region reduced to homozygosity in small-cell lung cancer (SCLC), and its expression has been reported to be reduced or undetectable in SCLC cell lines and tumors. The amino acid sequence of human aminoacylase-1 is highly homologous to the porcine counterpart, and this enzyme is the first member of a new family of zinc-binding enzymes. Mutations in ACY1 cause aminoacylase-1 deficiency, a metabolic disorder characterized by central nervous system defects and increased urinary excretion of N-acetylated amino acids. Alternative splicing of this gene results in multiple transcript variants. Read-through transcription also exists between ACY1 and the upstream ABHD14A (abhydrolase domain containing 14A) gene, as represented in GeneID: 100526760. A related pseudogene has been identified on chromosome 18. Application Notes: Optimal working dilutions should be determined experimentally by the investigator. Suggested starting dilutions are as follows: WB (1:500-1:2000), ELISA (1:5000-1:20000). Storage Buffer: PBS, pH 7.4, containing 0.02% Sodium Azide as preservative and 50% Glycerol.
    [Show full text]
  • Associated Gene
    Natural human genetic variation determines basal and inducible expression of PM20D1, an obesity- associated gene Kiara K. Bensona,b,c, Wenxiang Hua,b,c, Angela H. Wellera,b,c, Alexis H. Bennetta,b,c,1, Eric R. Chena,b,c,2, Sumeet A. Khetarpala,b,c,3, Satoshi Yoshinoa,b,c,4, William P. Boned,e,f, Lin Wangg,h, Joshua D. Rabinowitzg,h, Benjamin F. Voightd,e,f, and Raymond E. Soccioa,b,c,5 aDepartment of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104; bDivision of Endocrinology, Diabetes, and Metabolism, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104; cInstitute for Diabetes, Obesity, and Metabolism, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104; dDepartment of Genetics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104; eDepartment of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104; fInstitute of Translational Medicine and Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104; gDepartment of Chemistry, Princeton University, Princeton, NJ 08544; and hLewis Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544 Edited by Bruce M. Spiegelman, Dana-Farber Cancer Institute/Harvard Medical School, Boston, MA, and approved September 26, 2019 (received for review August 2, 2019) PM20D1 is a candidate thermogenic enzyme in mouse fat, with its without UCP1 to stimulate energy expenditure. NAA analogs are expression cold-induced and enriched in brown versus white adipo- thus in the early stages of pharmacological development (8). cytes. Thiazolidinedione (TZD) antidiabetic drugs, which activate the PM20D1 has also emerged from human genetic studies, most peroxisome proliferator-activated receptor-γ (PPARγ) nuclear recep- notably in neurodegenerative diseases.
    [Show full text]
  • ACY1 Rabbit Pab
    Leader in Biomolecular Solutions for Life Science ACY1 Rabbit pAb Catalog No.: A6351 Basic Information Background Catalog No. This gene encodes a cytosolic, homodimeric, zinc-binding enzyme that catalyzes the A6351 hydrolysis of acylated L-amino acids to L-amino acids and an acyl group, and has been postulated to function in the catabolism and salvage of acylated amino acids. This gene Observed MW is located on chromosome 3p21.1, a region reduced to homozygosity in small-cell lung 46kDa cancer (SCLC), and its expression has been reported to be reduced or undetectable in SCLC cell lines and tumors. The amino acid sequence of human aminoacylase-1 is highly Calculated MW homologous to the porcine counterpart, and this enzyme is the first member of a new 37kDa/38kDa/42kDa/45kDa family of zinc-binding enzymes. Mutations in this gene cause aminoacylase-1 deficiency, a metabolic disorder characterized by central nervous system defects and Category increased urinary excretion of N-acetylated amino acids. Alternative splicing of this gene results in multiple transcript variants. Read-through transcription also exists between Primary antibody this gene and the upstream ABHD14A (abhydrolase domain containing 14A) gene, as represented in GeneID:100526760. A related pseudogene has been identified on Applications chromosome 18. WB, IF Cross-Reactivity Human, Mouse, Rat Recommended Dilutions Immunogen Information WB 1:500 - 1:2000 Gene ID Swiss Prot 95 Q03154 IF 1:10 - 1:100 Immunogen Recombinant fusion protein containing a sequence corresponding to amino acids 1-408 of human ACY1 (NP_001185824.1). Synonyms ACY1;ACY-1;ACY1D;HEL-S-5 Contact Product Information www.abclonal.com Source Isotype Purification Rabbit IgG Affinity purification Storage Store at -20℃.
    [Show full text]