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Anti ATF6 alpha monoclonal antibody [Clone : 1-7]


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Background

ATF6 (activating transcription factor 6) is an endoplasmic reticulum (ER) membrane-bound transcription factor activated in response to ER stress. When unfolded proteins accumulate in the ER, ATF6 is cleaved by regulated intramembrane proteolysis. The resulting amino-terminal fragment translocates to the nucleus and activates transcription by binding to ER stress-response elements present in the promoter regions of ER stress-inducible genes including those encoding ER chaperones and components of ER-associated degradation. ATF6 consists of two closely related factors, ATF6a and ATF6b, in mammals. ATF6a but not ATF6b plays a pivotal role in transcriptional control.

Application

1) Western blotting
2) Immunoprecipitation (IP)
#  This antibody does not work for immunofluorescence analyses.
Data Link : Swiss-Prot P18850

Western blot analysis of human cell extracts using this antibody

Fig.1 Western blot analysis of human cell extracts using this antibody:Conversion of pATF6a(P) to pATF6a(N) in DTT- or tunicamycin-treated cells.
Tm: 2 g/ml tunicamycin (inhibitor of N-glycosylation).
DTT: 1mM dithiothreitol (reducing reagent).
The asterisk denotes an unglycosylated form of pATF6a (P).
ATF6a is constitutively expressed as pATF6a (P) (~90-kDa protein), and converted to pATF6a (N) (>50-kDa protein) in ER-stressed cells.

BAM_73500_2.png
Fig.2 IP-Western blot analysis of human cell extracts using this antibody.
ATF6a was detected by Western blot (Input; lanes 1, 2, 7, and 8) using this antibody (No.1-7). After immunoprecipitation (IP) with non-related IgG (IP; lanes 3, 4, 9, and 10) or this antibody (No. 1-7) (IP; lanes 5, 6, 11, and 12), samples were subjected to SDS-PAGE and analyzed by Western blot using this antibody (# 1-7) and anti-mouse IgG antibody (light chain specific).
Detection of pATF6a (P) and pATF6a (N) is better in IP-Western blotting than Western blotting.

<Reference>
1)   Hai T, et al., (1989) "Transcription factor ATF cDNA clones: an extensive family of leucine zipper proteins able to selectively form DNA-binding heterodimers." Genes Dev 3: 2083-2090 [PMID: 2516827]
2)   Haze K, et al., (1999) "Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress". Mol Biol Cell 10: 3787-3799 [PMID: 10564271]
3)   Yamamoto K. et al., (2007) "Transcriptional induction of mammalian ER quality control proteins is mediated by single or combined action of ATF6 and XBP1". Dev. Cell 13: 365-376[PMID: 17765680]

Product List

Product Name Cat# Quantity Price

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100 UG ¥42,100
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Anti ATF2, Human (Mouse) Unlabeled DataSheet

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Anti ATF3, Human (Mouse) Unlabeled DataSheet

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