Monoclonal Antibody to ATF6 Alpha

SM7007P OriGene Technologies Inc. OriGene EU

Acris Antibodies GmbH 9620 Medical Center Drive, Ste 200 Schillerstr. 5 Rockville, MD 20850 32052 Herford UNITED STATES GERMANY Phone: +1-858-888-7900 Phone: +49-5221-34606-0 Fax: +1-858-888-7904 Fax: +49-5221-34606-11 [email protected] [email protected]

Monoclonal Antibody to ATF6 alpha (full length + Cleaved Forms) - Purified Alternate names: ATF-6, Activating transcription factor 6 alpha, Cyclic AMP-dependent transcription factor ATF-6 alpha, cAMP-dependent transcription factor ATF-6 alpha Catalog No.: SM7007P Quantity: 0.1 mg Concentration: 0.5 mg/ml Background: ATF6 is a constitutively expressed, endoplasmic reticulum (ER) membrane-anchored transcription factor. ATF6 is a key transcriptional activator of the unfolded protein response (UPR), which allows mammalian cells to maintain cellular homeostasis when they are subjected to environmental and physiological stresses that target the ER (reviewed in Shen and Prywes, and Groenendyk and Michalak). The C-terminus of ATF6 is located in the ER lumen and its N-terminal DNA binding domain faces the cytosol. AFT6 plays a key role in the ER stress response by transmitting the ER stress signal across the ER membrane into the nucleus. The induction of new gene expression by ATF6 is an important aspect of the ER stress response. In response to certain stress conditions, ATF6 translocates from the ER to the Golgi. The 90 kDa full-length ATF6 is processed within the Golgi to its active 50 kDa form through sequential cleavage by site-1 and site-2 proteases (S1P and S2P). Proteolytic activation of ATF6 in the ER stress response is a mechanism to regulate membrane-bound factors, and is referred to as regulated intramembrane proteolysis. The N-terminal active ATF6 translocates to the nucleus where it binds to ER stress-response elements in ER stress- response genes (ERSRGs). ATF6 is a potent transcriptional activator of ERSRGs. The fully glycosylated form of ATF6, a 670 amino acid protein, exhibits an electrophoretic mobility of ~90 kDa in denaturing SDS-gels, in part because of the glycosylated modifications. ATF6 has 3 consensus sites for N-linked glycosylation and exists constitutively as a glycosylated protein. Differentially glycosylated ATF6 forms may result from mutations or experimental treatment (Shen and Prywes, 2005). Uniprot ID: P18850 NCBI: NP_031374.2 GeneID: 22926 Host / Isotype: Mouse / IgG1 Recommended SM10P (for use in human samples), SM20P (for use in rat samples), AM03095PU-N Isotype Controls: Clone: 70B1413.1 Immunogen: Partial protein containing amino acids 1-273 of Human ATF6.

For research and in vitro use only. Not for diagnostic or therapeutic work. MS/20140929 Material Safety Datasheets are available at www.acris-antibodies.com or on request. Acris Antibodies is now part of the OriGene family. Learn more at www.origene.com 1 / 4 SM7007P: Monoclonal Antibody to ATF6 alpha (full length + Cleaved Forms) - Purified Format: State: Liquid purified Ig fraction Purification: Protein G Chromatography Buffer System: PBS Preservatives: 0.05% Sodium Azide Stabilizers: 0.05% BSA Applications: Western blot analysis: 1-5 µg/ml. Recommended Positive Control: ATF6 transfected cell Lysates is recommended as useful Western Blot Postive Control to run in parallel with your experimental samples. Immunocytochemistry. Immunofluorescence: For Immunofluorescence microscopy, cells were fixed in methanol at -20°C (See Thomas et al, 2005 and Kikuchi et al, 2006). Thomas et al used Immunofluorescence to identify ATF6 with SM7007P in the nucleus. Immunohistochemistry (Paraffin): See van Kollenburg et al (2006) for details. Immunohistochemistry (Frozen): See Zhu et al, 2008 for details. ChIP: See Donati et al, 2006 and Renna et al, 2007 for details. Immunoprecipitation: See Hong et al, 2004 for details. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user. Specificity: The ATF6 antibody SM7007P has been cited to recognize both full-length and cleaved forms of ATF6. The active/cleaved 50 kDa nuclear form of ATF6 using Cat.-No SM7007P has been found to be strongly expressed in certain tumor cell lines derived from B cell lymphoma (DEL), primary effusion lymphoma [BC-3 (ATCC CRL-2277), PEL-SY, HBL-6], lymphoblastic leukemia (DS-1) and multiple myeloma (RPMI-8226, NCI-H929), (Jenner et al. 2003). Cleaved 60 and 36 kDa ATF6 forms have also been described in the nucleus (Mao et al, 2007). The ATF6 antibody is reported to be specific for ATF6 alpha, recognizing ATF6 alpha but not ATF6 beta (Bommiasamy et al, 2009). Species Reactivity: Tested: Human, Mouse, Rabbit, Rat, Hamster and Grouper (Fish). Storage: Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing. Shelf life: one year from despatch. General Readings: 1. Thomas M, Yu Z, Dadgar N, Varambally S, Yu J, Chinnaiyan AM, et al. The unfolded protein response modulates toxicity of the expanded glutamine androgen receptor. J Biol Chem. 2005 Jun 3;280(22):21264-71. Epub 2005 Mar 30. PubMed PMID: 15799970. 2. van Kollenburg B, van Dijk J, Garbern J, Thomas AA, Scheper GC, Powers JM, et al. Glia- specific activation of all pathways of the unfolded protein response in vanishing white matter disease. J Neuropathol Exp Neurol. 2006 Jul;65(7):707-15. PubMed PMID: 16825957. 3. Jenner RG, Maillard K, Cattini N, Weiss RA, Boshoff C, Wooster R, et al. Kaposi's sarcoma- associated herpesvirus-infected primary effusion lymphoma has a plasma cell gene expression profile. Proc Natl Acad Sci U S A. 2003 Sep 2;100(18):10399-404. Epub 2003 Aug 18. PubMed PMID: 12925741. 4. Donati G, Imbriano C, Mantovani R. Dynamic recruitment of transcription factors and epigenetic changes on the ER stress response gene promoters. Nucleic Acids Res. 2006 Jun 6;34(10):3116-27. Print 2006. PubMed PMID: 16757577. 5. Hong M, Li M, Mao C, Lee AS. Endoplasmic reticulum stress triggers an acute proteasome- dependent degradation of ATF6. J Cell Biochem. 2004 Jul 1;92(4):723-32. PubMed PMID: 15211570. 6. Requirement of the p38 mitogen-activated protein kinase signalling pathway for the induction of the 78kDa glucose-regulated protein / immunoglobulin heavy-chain binding protein by azetidine stress: activating transcription factor 6 as a target for stress-induced

Western blot analysis of SM7007P ATF6 antibody. Lane 1: 293 cells transfected with full-length ATF6. Lane 2: 293 cells transfected with partial length ATF6 (amino acids 1-373). Lane 3: Untransfected 293 cells. Western blots were probed with 4 ug/ml of the monoclonal SM7007P ATF6 antibody followed by an HRP-conjugated second step and visualized with PicoTect Western Blot Chemiluminescence Substrate. Film was exposed for 1 min. The top arrow corresponds to the ~90 kDa form of ATF6 described as full-length in the literature. The human full-length and partial length ATF6 plasmids are described in Luo and Lee (2002).