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Technical Note: Colorimetric Protein Assays Assays for Determining Protein Concentration
Technical Note: Colorimetric Protein Assays Assays for Determining Protein Concentration Contents speed. No colorimetric assay is as simple and quick as direct UV measurements at 280 nm for Introduction ....................................................... 1 the determination of protein concentration. Assay Choice ..................................................... 1 However measurements at 280 nm rely on the Buffer Composition ........................................ 1 protein containing aromatic amino acids such as Sample Composition ...................................... 2 tyrosine (Y), phenylalanine (F), and, or tryptophan Standard Selection ......................................... 2 (W). Not all proteins contain these amino acids, Assays ................................................................ 2 and the relative proportions of theses amino acids Biuret, BCA, and Lowry Assays ..................... 3 differ between proteins. Furthermore, if nucleic Bradford Assay............................................... 3 acids are present in the sample, they would also Troubleshooting ............................................. 4 absorb light at 280 nm, further compromising FAQ’s .................................................................. 5 accuracy. Therefore, the sensitivity achieved Bibliography ...................................................... 5 without prior knowledge of the protein of interest’s Contact Us ......................................................... 6 absorbance maxima (λmax) and attenuation coefficient -
Phenolphthalein, 0.5% in 50% Ethanol Safety Data Sheet According to Federal Register / Vol
Phenolphthalein, 0.5% in 50% Ethanol Safety Data Sheet according to Federal Register / Vol. 77, No. 58 / Monday, March 26, 2012 / Rules and Regulations Issue date: 12/17/2013 Revision date: 07/21/2020 Supersedes: 01/23/2018 Version: 1.5 SECTION 1: Identification 1.1. Identification Product form : Mixtures Product name : Phenolphthalein, 0.5% in 50% Ethanol Product code : LC18200 1.2. Recommended use and restrictions on use Use of the substance/mixture : For laboratory and manufacturing use only. Recommended use : Laboratory chemicals Restrictions on use : Not for food, drug or household use 1.3. Supplier LabChem, Inc. 1010 Jackson's Pointe Ct. Zelienople, PA 16063 - USA T 412-826-5230 - F 724-473-0647 [email protected] - www.labchem.com 1.4. Emergency telephone number Emergency number : CHEMTREC: 1-800-424-9300 or +1-703-741-5970 SECTION 2: Hazard(s) identification 2.1. Classification of the substance or mixture GHS US classification Flammable liquids Category 3 H226 Flammable liquid and vapor Carcinogenicity Category 1A H350 May cause cancer Reproductive toxicity Category 2 H361 Suspected of damaging the unborn child. (oral) Specific target organ toxicity (single exposure) Category 1 H370 Causes damage to organs (central nervous system, optic nerve) (oral, Dermal) Full text of H statements : see section 16 2.2. GHS Label elements, including precautionary statements GHS US labeling Hazard pictograms (GHS US) : Signal word (GHS US) : Danger Hazard statements (GHS US) : H226 - Flammable liquid and vapor H350 - May cause cancer H361 - Suspected of damaging the unborn child. (oral) H370 - Causes damage to organs (central nervous system, optic nerve) (oral, Dermal) Precautionary statements (GHS US) : P201 - Obtain special instructions before use. -
The Action of the Phosphatases of Human Brain on Lipid Phosphate Esters by K
J Neurol Neurosurg Psychiatry: first published as 10.1136/jnnp.19.1.12 on 1 February 1956. Downloaded from J. Neurol. Neurosurg. Psychiat., 1956, 19, 12 THE ACTION OF THE PHOSPHATASES OF HUMAN BRAIN ON LIPID PHOSPHATE ESTERS BY K. P. STRICKLAND*, R. H. S. THOMPSON, and G. R. WEBSTER From the Department of Chemical Pathology, Guy's Hospital Medical School, London, Much work, using both histochemical and therefore to study the action of the phosphatases in standard biochemical techniques, has been carried human brain on the " lipid phosphate esters out on the phosphatases of peripheral nerve. It is i.e., on the various monophosphate esters that occur known that this tissue contains both alkaline in the sphingomyelins, cephalins, and lecithins. In (Landow, Kabat, and Newman, 1942) and acid addition to ox- and 3-glycerophosphate we have phosphatases (Wolf, Kabat, and Newman, 1943), therefore used phosphoryl choline, phosphoryl and the changes in the levels of these enzymes in ethanolamine, phosphoryl serine, and inositol nerves undergoing Wallerian degeneration following monophosphate as substrates for the phospho- transection have been studied by several groups of monoesterases, and have measured their rates guest. Protected by copyright. of investigators (see Hollinger, Rossiter, and Upmalis, hydrolysis by brain preparations over the pH range 1952). 4*5 to 100. Phosphatase activity in brain was first demon- Plimmer and Burch (1937) had earlier reported strated by Kay (1928), and in 1934 Edlbacher, that phosphoryl choline and phosphoryl ethanol- Goldschmidt, and Schiiippi, using ox brain, showed amine are hydrolysed by the phosphatases of bone, that both acid and alkaline phosphatases are kidney, and intestine, but thepH at which the hydro- present in this tissue. -
Student Safety Sheets Dyes, Stains & Indicators
Student safety sheets 70 Dyes, stains & indicators Substance Hazard Comment Solid dyes, stains & indicators including: DANGER: May include one or more of the following Acridine orange, Congo Red (Direct dye 28), Crystal violet statements: fatal/toxic if swallowed/in contact (methyl violet, Gentian Violet, Gram’s stain), Ethidium TOXIC HEALTH with skin/ if inhaled; causes severe skin burns & bromide, Malachite green (solvent green 1), Methyl eye damage/ serious eye damage; may cause orange, Nigrosin, Phenolphthalein, Rosaniline, Safranin allergy or asthma symptoms or breathing CORR. IRRIT. difficulties if inhaled; may cause genetic defects/ cancer/damage fertility or the unborn child; causes damages to organs/through prolonged or ENVIRONMENT repeated exposure. Solid dyes, stains & indicators including Alizarin (1,2- WARNING: May include one or more of the dihydroxyanthraquinone), Alizarin Red S, Aluminon (tri- following statements: harmful if swallowed/in ammonium aurine tricarboxylate), Aniline Blue (cotton / contact with skin/if inhaled; causes skin/serious spirit blue), Brilliant yellow, Cresol Red, DCPIP (2,6-dichl- eye irritation; may cause allergic skin reaction; orophenolindophenol, phenolindo-2,6-dichlorophenol, HEALTH suspected of causing genetic PIDCP), Direct Red 23, Disperse Yellow 7, Dithizone (di- defects/cancer/damaging fertility or the unborn phenylthiocarbazone), Eosin (Eosin Y), Eriochrome Black T child; may cause damage to organs/respiratory (Solochrome black), Fluorescein (& disodium salt), Haem- HARMFUL irritation/drowsiness or dizziness/damage to atoxylin, HHSNNA (Patton & Reeder’s indicator), Indigo, organs through prolonged or repeated exposure. Magenta (basic Fuchsin), May-Grunwald stain, Methyl- ene blue, Methyl green, Orcein, Phenol Red, Procion ENVIRON. dyes, Pyronin, Resazurin, Sudan I/II/IV dyes, Sudan black (Solvent Black 3), Thymol blue, Xylene cyanol FF Solid dyes, stains & indicators including Some dyes may contain hazardous impurities and Acid blue 40, Blue dextran, Bromocresol green, many have not been well researched. -
Phenolphthalein Is Pink in Base Acid–Base Indicators SCIENTIFIC
Phenolphthalein Is Pink in Base Acid–Base Indicators SCIENTIFIC Introduction Phenolphthalein is a large organic molecule used as an acid–base indicator. Phenolphthalein turns a bright red color as its solution becomes basic. In a strongly basic solution, this red color fades to colorless. Concepts • Acid–base indicators • Chemical equilibrium Background Phenolphthalein has the colorless structure shown in Figure 1 when the solution pH <8. As the solution becomes basic and the pH increases (pH 8–10), the phenolphthalein molecule (abbreviated H2P) loses two hydrogen ions to form the red- violet dianion (abbreviated P2–) shown in Figure 2. At a high pH, the P2– ions reacts with hydroxide ions to form the colorless POH3– ion. HO – – O –O O O OH O O– C C C – – + OH C O CO– 2 CO2 OH – CO2 O P2– Red 2– POH3– Colorless3– Figure 1. H2P is colorless. Figure 2. P is red. Figure 3. POH is colorless. 2– The colorless-to-red transition of H2P to P (Equation 1) is very rapid and the red color develops instantly when the pH reaches its transition range (pH 8–10). If the concentration of hydroxide ions remains high, the red P2– dianion will slowly combine with hydroxide ions to form a third species, POH3– (Equation 2), which is again colorless. The rate of this second reaction is much slower than the first and depends on the concentration of phenolphthalein and hydroxide ions. Thus, the color of the red P2– species will gradually fade in a basic solution. fast 2– + H2P → P + 2H Equation 1 Colorless Red slow P2– + OH– → POH3– Equation 2 Red Colorless Materials (for each demonstration) Hydrochloric acid solution, HCl, 3 M, 10 mL Beral-type pipet, disposable Phenolphthalein solution, 0.5%, 3mL Test tubes, borosilicate, 16 5 100mm, 3 Sodium hydroxide pellets, NaOH, 2 Test tube rack Sodium hydroxide solution, NaOH, 3 M, 5 mL Wash bottle Safety Precautions Hydrochloric acid solution is toxic and corrosive to eyes and skin tissue. -
Thermostable and Long-Circulating Albumin-Conjugated Arthrobacter Globiformis Urate Oxidase
pharmaceutics Article Thermostable and Long-Circulating Albumin-Conjugated Arthrobacter globiformis Urate Oxidase Byungseop Yang and Inchan Kwon * School of Materials Science and Engineering, Gwangju Institute of Science and Technology (GIST), Gwangju 61005, Korea; [email protected] * Correspondence: [email protected]; Tel.: +82-62-715-2312 Abstract: Urate oxidase derived from Aspergillus flavus has been investigated as a treatment for tumor lysis syndrome, hyperuricemia, and gout. However, its long-term use is limited owing to potential immunogenicity, low thermostability, and short circulation time in vivo. Recently, urate oxidase isolated from Arthrobacter globiformis (AgUox) has been reported to be thermostable and less immunogenic than the Aspergillus-derived urate oxidase. Conjugation of human serum albumin (HSA) to therapeutic proteins has become a promising strategy to prolong circulation time in vivo. To develop a thermostable and long-circulating urate oxidase, we investigated the site-specific conjugation of HSA to AgUox based on site-specific incorporation of a clickable non-natural amino acid (frTet) and an inverse electron demand Diels–Alder reaction. We selected 14 sites for frTet incorporation using the ROSETTA design, a computational stability prediction program, among which AgUox containing frTet at position 196 (Ag12) exhibited enzymatic activity and thermostability comparable to those of wild-type AgUox. Furthermore, Ag12 exhibited a high HSA conjugation yield without compromising the enzymatic activity, generating well-defined HSA-conjugated AgUox (Ag12-HSA). In mice, the serum half-life of Ag12-HSA was approximately 29 h, which was roughly Citation: Yang, B.; Kwon, I. 17-fold longer than that of wild-type AgUox. Altogether, this novel formulated AgUox may hold Thermostable and Long-Circulating enhanced therapeutic efficacy for several diseases. -
UDP-N-Acetylglucosamine Transporter and UDP-Galactose Transporter
View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector FEBS Letters 586 (2012) 4082–4087 journal homepage: www.FEBSLetters.org UDP-N-acetylglucosamine transporter and UDP-galactose transporter form heterologous complexes in the Golgi membrane ⇑ Dorota Maszczak-Seneczko a, Paulina Sosicka a, Michał Majkowski b, Teresa Olczak a, Mariusz Olczak a, a Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Wroclaw, Poland b Laboratory of Cytobiochemistry, Faculty of Biotechnology, University of Wroclaw, Wroclaw, Poland article info abstract Article history: UDP-galactose transporter (UGT; SLC35A2) and UDP-N-acetylglucosamine transporter (NGT; Received 6 August 2012 SLC35A3) are evolutionarily related. We hypothesize that their role in glycosylation may be coupled Revised 2 October 2012 through heterologous complex formation. Coimmunoprecipitation analysis and FLIM–FRET mea- Accepted 8 October 2012 surements performed on living cells showed that NGT and UGT form complexes when overexpressed Available online 23 October 2012 in MDCK-RCAr cells. We also postulate that the interaction of NGT and UGT may explain the dual Edited by Judit Ovádi localization of UGT2. For the first time we demonstrated in vivo homodimerization of the NGT nucle- otide sugar transporter. In conclusion, we suggest that NGT and UGT function in glycosylation is combined via their mutual interaction. Keywords: UDP-N-acetylglucosamine transporter UDP-galactose transporter Structured summary -
Step-By-Step Guide to Better Laboratory Management Practices
Step-by-Step Guide to Better Laboratory Management Practices Prepared by The Washington State Department of Ecology Hazardous Waste and Toxics Reduction Program Publication No. 97- 431 Revised January 2003 Printed on recycled paper For additional copies of this document, contact: Department of Ecology Publications Distribution Center PO Box 47600 Olympia, WA 98504-7600 (360) 407-7472 or 1 (800) 633-7585 or contact your regional office: Department of Ecology’s Regional Offices (425) 649-7000 (509) 575-2490 (509) 329-3400 (360) 407-6300 The Department of Ecology is an equal opportunity agency and does not discriminate on the basis of race, creed, color, disability, age, religion, national origin, sex, marital status, disabled veteran’s status, Vietnam Era veteran’s status or sexual orientation. If you have special accommodation needs, or require this document in an alternate format, contact the Hazardous Waste and Toxics Reduction Program at (360)407-6700 (voice) or 711 or (800) 833-6388 (TTY). Table of Contents Introduction ....................................................................................................................................iii Section 1 Laboratory Hazardous Waste Management ...........................................................1 Designating Dangerous Waste................................................................................................1 Counting Wastes .......................................................................................................................8 Treatment by Generator...........................................................................................................12 -
Disclosing the Essentiality of Ribose-5-Phosphate Isomerase B In
www.nature.com/scientificreports OPEN Disclosing the essentiality of ribose-5-phosphate isomerase B in Trypanosomatids Received: 04 January 2016 Joana Faria1,2, Inês Loureiro1,2, Nuno Santarém1,2, Pedro Cecílio1,2, Sandra Macedo-Ribeiro2,3, Accepted: 10 May 2016 Joana Tavares1,2,* & Anabela Cordeiro-da-Silva1,2,4,* Published: 27 May 2016 Ribose-5-phosphate isomerase (RPI) belongs to the non-oxidative branch of the pentose phosphate pathway, catalysing the inter-conversion of D-ribose-5-phosphate and D-ribulose-5-phosphate. Trypanosomatids encode a type B RPI, whereas humans have a structurally unrelated type A, making RPIB worthy of exploration as a potential drug target. Null mutant generation in Leishmania infantum was only possible when an episomal copy of RPIB gene was provided, and the latter was retained both in vitro and in vivo in the absence of drug pressure. This suggests the gene is essential for parasite survival. Importantly, the inability to remove the second allele of RPIB gene in sKO mutants complemented with an episomal copy of RPIB carrying a mutation that abolishes isomerase activity suggests the essentiality is due to its metabolic function. In vitro, sKO promastigotes exhibited no defect in growth, metacyclogenesis or macrophage infection, however, an impairment in intracellular amastigotes’ replication was observed. Additionally, mice infected with sKO mutants rescued by RPIB complementation had a reduced parasite burden in the liver. Likewise, Trypanosoma brucei is resistant to complete RPIB gene removal and mice infected with sKO mutants showed prolonged survival upon infection. Taken together our results genetically validate RPIB as a potential drug target in trypanosomatids. -
IER5 Is Involved in DNA Double-Strand Breaks Repair In
Int. J. Med. Sci. 2017, Vol. 14 1292 Ivyspring International Publisher International Journal of Medical Sciences 2017; 14(12): 1292-1300. doi: 10.7150/ijms.21510 Research Paper IER5 is involved in DNA Double-Strand Breaks Repair in Association with PAPR1 in Hela Cells Xin-Ping Yu1, Yu-Mei Wu1, Yang Liu1, Ming Tian1, Jian-Dong Wang1, Ku-Ke Ding2, Teng Ma3, Ping-Kun Zhou3 1. Department of Gynecologic Oncology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, 100006, China; 2. National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing ,100088, China; 3. Department of Radiation Toxicology and Oncology, Beijing Key Laboratory for Radiobiology, Beijing Institute of Radiation Medicine, Beijing, 100850, China. Corresponding author: Yu-Mei Wu, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, 17 Qi-he-lou St, Dongcheng District, Beijing 100006, China. E-mail: [email protected] © Ivyspring International Publisher. This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license (https://creativecommons.org/licenses/by-nc/4.0/). See http://ivyspring.com/terms for full terms and conditions. Received: 2017.06.17; Accepted: 2017.09.01; Published: 2017.09.30 Abstract The immediate early response gene 5 (IER5) is a radiation response gene induced in a dose-independent manner, and has been suggested to be a molecular biomarker for biodosimetry purposes upon radiation exposure. Here, we investigated the function of IER5 in DNA damage response and repair. We found that interference on IER5 expression significantly decreased the efficiency of repair of DNA double-strand breaks induced by ionizing radiations in Hela cells. -
Evaluation of Signaling Pathways Profiling in Human Dermal
toxins Article Evaluation of Signaling Pathways Profiling in Human Dermal Endothelial Cells Treated by Snake Venom Cysteine-Rich Secretory Proteins (svCRiSPs) from North American Snakes Using Reverse Phase Protein Array (RPPA) Montamas Suntravat 1,2,* , Oscar Sanchez 1, Armando Reyes 1, Abcde Cirilo 1, Jack S. Ocheltree 1, Jacob A. Galan 1,2, Emelyn Salazar 1 , Peter Davies 3 and Elda E. Sanchez 1,2 1 National Natural Toxins Research Center (NNTRC), Texas A&M University-Kingsville, MSC 224, 975 West Avenue B, Kingsville, TX 78363, USA; [email protected] (O.S.); [email protected] (A.R.); [email protected] (A.C.); [email protected] (J.S.O.); [email protected] (J.A.G.); [email protected] (E.S.); [email protected] (E.E.S.) 2 Department of Chemistry, Texas A&M University-Kingsville, MSC 161, Kingsville, TX 78363, USA 3 Institute of Biosciences and Technology, Texas A&M University, Houston, TX 77843, USA; [email protected] * Correspondence: [email protected] Abstract: Cysteine-Rich Secretory Proteins (CRiSPs) are typically found in many snake venoms; however, the role that these toxins play in the pathophysiology of snakebites is still unclear. Herein, we compared the effects of snake venom CRiSPs (svCRiSPs) from the most medically important Citation: Suntravat, M.; Sanchez, O.; species of North American snakes on endothelial cell permeability and vascular permeability. We Reyes, A.; Cirilo, A.; Ocheltree, J.S.; used reverse phase protein array (RPPA) to identify key signaling molecules on human dermal Galan, J.A.; Salazar, E.; Davies, P.; lymphatic (HDLECs) and blood (HDBECs) endothelial cells treated with svCRiSPs. -
MATERIAL SAFETY DATA SHEET Phenolphthalein
MATERIAL SAFETY DATA SHEET Phenolphthalein Section 1 Chemical Product and Company Identification MSDS Name: Phenolphthalein Catalog 147710000, 147711000, 147715000, 417180000, 417180025, 417181000, 41718 Numbers: 5000 Synonyms: Company Identification: Acros Organics BVBA Janssen Pharmaceuticalaan 3a 2440 Geel, Belgium Company Identification: (USA) Acros Organics One Reagent Lane Fair Lawn, NJ 07410 For information in the US, call: 800ACROS01 For information in Europe, call: +32 14 57 52 11 Emergency Number, Europe: +32 14 57 52 99 Emergency Number US: 2017967100 CHEMTREC Phone Number, US: 8004249300 CHEMTREC Phone Number, Europe: 7035273887 Section 2 Composition, Information on Ingredients CAS# Chemical Name: % EINECS# 77098 Phenolphthalein 2010047 Hazard Symbols: XN Risk Phrases: 22 40 Section 3 Hazards Identification EMERGENCY OVERVIEW Harmful if swallowed. Limited evidence of a carcinogenic effect. Potential Health Effects Eye: May cause eye irritation. Skin: May cause skin irritation. May be harmful if absorbed through the skin. Ingestion: Harmful if swallowed. May cause irritation of the digestive tract. Ingestion may cause fever, blood pressure increase and other unspecified vascular effects. Inhalation: May cause respiratory tract irritation. May be harmful if inhaled. Chronic: Limited evidence of a carcinogenic effect. Section 4 First Aid Measures Eyes: Flush eyes with plenty of water for at least 15 minutes, occasionally lifting the upper and lower eyelids. Get medical aid. Skin: Get medical aid. Flush skin with plenty of water for at least 15 minutes while removing contaminated clothing and shoes. Ingestion: Get medical aid. Wash mouth out with water.