ATF6 Monoclonal antibody

• Featured Product
• KD/KO Validated

ATF6 Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat and More (1)

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

30

CloneNo.

3B7E4

Cat no : 66563-1-Ig

Synonyms

ATF6-alpha, ATF6A, ATF6 alpha, ATF 6, activating transcription factor 6

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Validation Data Gallery

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  WB analysis using 66563-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66563-1-Ig (ATF6 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis using 66563-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66563-1-Ig (ATF6 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 66563-1-Ig

  WB result of ATF6 antibody (66563-1-Ig; 1:30000; incubated at room temperature for 1.5 hours) with sh-Control and sh-ATF6 transfected HEK-293 cells.

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  WB analysis of HeLa using 66563-1-Ig

  WB result of ATF6 antibody (66563-1-Ig; 1:30000; incubated at room temperature for 1.5 hours) with sh-Control and sh-ATF6 transfected HeLa cells.

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  WB analysis of HeLa using 66563-1-Ig

  HeLa cells were subjected to SDS PAGE followed by western blot with 66563-1-Ig (ATF6 antibody) at dilution of 1:5000 incubated at 4 degree celsius over night.

• 

  WB analysis using 66563-1-Ig

  Various cells were subjected to SDS PAGE followed by western blot with 66563-1-Ig (ATF6 antibody) at dilution of 1:5000 incubated at 4 degree celsius over night.

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  WB analysis of HSC-T6 using 66563-1-Ig

  HSC-T6 cells were subjected to SDS PAGE followed by western blot with 66563-1-Ig (ATF6 antibody) at dilution of 1:5000 incubated at 4 degree celsius over night.

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  WB analysis of NIH/3T3 using 66563-1-Ig

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 66563-1-Ig (ATF6 antibody) at dilution of 1:5000 incubated at 4 degree celsius over night.

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  WB analysis of RAW 264.7 using 66563-1-Ig

  RAW 264.7 cells were subjected to SDS PAGE followed by western blot with 66563-1-Ig (ATF6 antibody) at dilution of 1:5000 incubated at 4 degree celsius over night.

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  WB analysis of 4T1 using 66563-1-Ig

  4T1 cells were subjected to SDS PAGE followed by western blot with 66563-1-Ig (ATF6 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IHC staining of human cervical cancer using 66563-1-Ig

  Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue slide using 66563-1-Ig (ATF6 antibody) at dilution of 1:500 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66563-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66563-1-Ig (ATF6 antibody) at dilution of 1:500 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66563-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66563-1-Ig (ATF6 antibody) at dilution of 1:500 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human cervical cancer using 66563-1-Ig

  Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue slide using 66563-1-Ig (ATF6 antibody) at dilution of 1:500 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 66563-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using ATF6 antibody (66563-1-Ig, Clone: 3B7E4 ) at dilution of 1:800 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (RGAM002), CL594-phalloidin (red).

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  FC experiment of HeLa using 66563-1-Ig

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human ATF6 (66563-1-Ig, Clone:3B7E4) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: U2OS cells, HeLa cells, HEK-293 cells, 4T1 cells, HSC-T6 cells, NIH/3T3 cells, RAW 264.7 cells, MCF-7 cells, Jurkat cells, K-562 cells
• Positive IHC detected in: human cervical cancer tissue, human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 2 publications below
• WB: See 27 publications below
• IHC: See 1 publications below
• IF: See 3 publications below

Product Information

66563-1-Ig targets ATF6 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, pig
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: ATF6 fusion protein Ag21456
• Full Name: activating transcription factor 6
• Calculated Molecular Weight: 75 kDa
• Observed Molecular Weight: 90-100 kDa
• GenBank Accession Number: BC014969
• Gene Symbol: ATF6
• Gene ID (NCBI): 22926
• RRID: AB_2881924
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Activating transcription factor 6 (ATF6) is a transcription factor that acts during endoplasmic reticulum stress by activating unfolded protein response target genes. Binds DNA on the 5'-CCAC[GA]-3'half of the ER stress response element (ERSE) (5'-CCAAT-N(9)-CCAC[GA]-3') and of ERSE II (5'-ATTGG-N-CCACG-3'). Binding to ERSE requires binding of NF-Y to ERSE. Could also be involved in activation of transcription by the serum response factor.During unfolded protein response an approximative 50 kDa fragment containing the cytoplasmic transcription factor domain is released by proteolysis. The cleavage seems to be performed sequentially by site-1 and site-2 proteases. 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 (PMID:15804611) (PMID:14699159). The antibody recognizes cleaved and fully glycosylated forms of ATF6.

Publications

Species	Application	Title
human	WB	Protein Cell; POST1/C12ORF49 regulates the SREBP pathway by promoting site-1 protease maturation.; Authors - Jian Xiao
mouse	WB	J Cell Biol; Antigen-derived peptides engage the ER stress sensor IRE1α to curb dendritic cell cross-presentation.; Authors - Ofer Guttman
human	WB	Cell Rep; CD44 correlates with longevity and enhances basal ATF6 activity and ER stress resistance; Authors - Masaki Takasugi
human	WB	Elife; Endoplasmic reticulum stress activates human IRE1α through reversible assembly of inactive dimers into small oligomers.; Authors - Vladislav Belyy
human	WB	Mol Oncol; The synthetic oleanane triterpenoid CDDO-2P-Im binds GRP78/BiP to induce unfolded protein response-mediated apoptosis in myeloma; Authors - George Luo
human	WB,IF	J Inflamm (Lond); Soluble epoxide hydrolase deficiency attenuates airway inflammation in COPD via IRE1α/JNK/AP-1 signaling pathway; Authors - Yue Yu