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ATF6 Mouse Monoclonal Antibody [8D3]

Usd: 350

Specification

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Catalog# EM1701-94

ATF6 Mouse Monoclonal Antibody [8D3]

Application

  • WB

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

ATF6 Mouse Monoclonal Antibody [8D3]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within Human ATF6 aa 1-670 / 670.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 75 kDa

Positive Control

HeLa cell lysate, Jurkat cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, HeLa, mouse kidney tissue lysate, rat brain tissue, human placenta tissue, mouse testis tissue, HepG2.

Conjugation

unconjugated

Clone Number

8D3

RRID

AB_3068817

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein G affinity purified.

Application Dilution

  • WB

  • 1:1000-1:5,000

  • IF-Cell

  • 1:100

Target

Function

Transmembrane glycoprotein of the endoplasmic reticulum that functions as a transcription activator and initiates the unfolded protein response (UPR) during endoplasmic reticulum stress. Cleaved upon ER stress, the N-terminal processed cyclic AMP-dependent transcription factor ATF-6 alpha translocates to the nucleus where it activates transcription of genes involved in the UPR. Binds DNA on the 5'-CCAC[GA]-3'half of the ER stress response element (ERSE) (5'-CCAAT-N9-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. May play a role in foveal development and cone function in the retina.

Background References

1. Kohl S et al. Mutations in the unfolded protein response regulator ATF6 cause the cone dysfunction disorder achromatopsia. Nat Genet 47:757-765 (2015).

2. Lynch J M et al. A thrombospondin-dependent pathway for a protective ER stress response. Cell 149:1257-1268 (2012).

Sequence Similarity

Belongs to the bZIP family. ATF subfamily.

Tissue Specificity

Ubiquitous.

Post-translational Modification

During unfolded protein response, a fragment of approximately 50 kDa containing the cytoplasmic transcription factor domain is released by proteolysis. The cleavage seems to be performed sequentially by site-1 and site-2 proteases.; N-glycosylated. The glycosylation status may serve as a sensor for ER homeostasis, resulting in ATF6 activation to trigger the unfolded protein response (UPR).; Phosphorylated in vitro by MAPK14/P38MAPK.

Subcellular Location

Endoplasmic reticulum. Nucleus.

UNIPROT

SwissProt: P18850 Human

SwissProt: F6VAN0 Mouse

SwissProt: G3V909 Rat

Synonyms

Activating transcription factor 6 alpha antibody

Activating transcription factor 6 antibody

ATF 6 antibody

ATF6 alpha antibody

ATF6 antibody

ATF6-alpha antibody

ATF6A antibody

ATF6A_HUMAN antibody

cAMP dependent transcription factor ATF 6 alpha antibody

cAMP-dependent transcription factor ATF-6 alpha antibody

Expand

Images

  • Western blot analysis of ATF6 on different lysates with Mouse anti-ATF6 antibody (<a href="/products/EM1701-94" style="font-weight: bold;text-decoration: underline;">EM1701-94</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: Jurkat cell lysate (20 µg/Lane)<br />Lane 3: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 4: PC-12 cell lysate (20 µg/Lane)<br />Lane 5: Mouse brain tissue lysate (40 µg/Lane)<br />Lane 6: Mouse kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 75 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 24 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1701-94" style="font-weight: bold;text-decoration: underline;">EM1701-94</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of ATF6 on different lysates with Mouse anti-ATF6 antibody (EM1701-94) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: Jurkat cell lysate (20 µg/Lane)
    Lane 3: RAW264.7 cell lysate (20 µg/Lane)
    Lane 4: PC-12 cell lysate (20 µg/Lane)
    Lane 5: Mouse brain tissue lysate (40 µg/Lane)
    Lane 6: Mouse kidney tissue lysate (40 µg/Lane)

    Predicted band size: 75 kDa
    Observed band size: 75 kDa

    Exposure time: 24 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-94) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling ATF6 with Mouse anti-ATF6 antibody (<a href="/products/EM1701-94" style="font-weight: bold;text-decoration: underline;">EM1701-94</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-ATF6 antibody (<a href="/products/EM1701-94" style="font-weight: bold;text-decoration: underline;">EM1701-94</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling ATF6 with Mouse anti-ATF6 antibody (EM1701-94) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-ATF6 antibody (EM1701-94) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of PC-12 cells labeling ATF6 with Mouse anti-ATF6 antibody (<a href="/products/EM1701-94" style="font-weight: bold;text-decoration: underline;">EM1701-94</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-ATF6 antibody (<a href="/products/EM1701-94" style="font-weight: bold;text-decoration: underline;">EM1701-94</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of PC-12 cells labeling ATF6 with Mouse anti-ATF6 antibody (EM1701-94) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-ATF6 antibody (EM1701-94) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of RAW264.7 cells labeling ATF6 with Mouse anti-ATF6 antibody (<a href="/products/EM1701-94" style="font-weight: bold;text-decoration: underline;">EM1701-94</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-ATF6 antibody (<a href="/products/EM1701-94" style="font-weight: bold;text-decoration: underline;">EM1701-94</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of RAW264.7 cells labeling ATF6 with Mouse anti-ATF6 antibody (EM1701-94) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-ATF6 antibody (EM1701-94) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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