Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Porcine

Applications

Validated:

Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all ATF6 Primary Antibodies »

Product Specifications

Immunogen

A partial recombinant protein made to an internal region of the human ATF6 protein (within residues 200-350). [Swiss-Prot P18850]

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 31235574).

Localization

Endoplasmic reticulum membrane, nucleus. Under ER stress the cleaved N-terminal cytoplasmic domain translocates into the nucleus.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

Novus Biologicals Rabbit ATF6 Antibody - BSA Free (NBP1-75478) is a polyclonal antibody validated for use in WB, ICC/IF and Simple Western. Anti-ATF6 Antibody: Cited in 6 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for ATF6 Antibody - BSA Free

Western Blot: ATF6 AntibodyBSA Free [NBP1-75478]

Western Blot: ATF6 AntibodyBSA Free [NBP1-75478]

Western Blot: ATF6 Antibody [NBP1-75478] - Total protein from HeLa cells treated with and without 1 mM DTT for 24 hours was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 1% BSA. The membrane was probed with 2.0 ug/ml anti-ATF6 in block buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the decrease in full length ATF6 (90 kDa, arrow) and increase in cleaved ATF6 (55 kDa, arrowhead) upon DTT treatment and the activation of the unfolded protein response pathway.
Simple Western: ATF6 AntibodyBSA Free [NBP1-75478]

Simple Western: ATF6 AntibodyBSA Free [NBP1-75478]

Simple Western: ATF6 Antibody [NBP1-75478] - Simple Western lane view shows a specific band for ATF6 in 1.0 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230kDa separation system.
Immunocytochemistry/ Immunofluorescence: ATF6 Antibody - BSA Free [NBP1-75478]

Immunocytochemistry/ Immunofluorescence: ATF6 Antibody - BSA Free [NBP1-75478]

Immunocytochemistry/Immunofluorescence: ATF6 Antibody [NBP1-75478] - HeLa cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-ATF6 at 10 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Western Blot: ATF6 AntibodyBSA Free [NBP1-75478]

Western Blot: ATF6 AntibodyBSA Free [NBP1-75478]

ATF6-Antibody-Western-Blot-NBP1-75478-img0007.jpg
Immunocytochemistry/ Immunofluorescence: ATF6 Antibody - BSA Free [NBP1-75478]

Immunocytochemistry/ Immunofluorescence: ATF6 Antibody - BSA Free [NBP1-75478]

Immunocytochemistry/Immunofluorescence: ATF6 Antibody [NBP1-75478] - Antibody was tested in HeLa cells with FITC (green). Nuclei were counterstained with Dapi (blue).

Applications for ATF6 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100

Simple Western

1:25

Western Blot

1:100-1:2000
Application Notes

This ATF6 antibody is useful for Immunocytochemistry/Immunofluorescence.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 1.0 mg/mL, separated by Size, antibody dilution of 1:25, apparent MW was 101 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 1 review rated 1 using NBP1-75478 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: ATF6

Endoplasmic reticulum stress results in protein folding irregularities and proteolytic abberations. ER stress is mediated in part by the activating transcription factor (ATF6). ATF6 is a signal transducer that prohibits phosphorylation of certain proteins, like the insulin receptor, and protects overactive responses to ER stress. This ultimately mediates apoptosis through mitochondrial cellular signaling. Influenza infections are known to affect ER stress in lung endothelial cells, partially mediated by ATF6 activity. This protein is also being investigated for potential optimization of recovery efforts, following traumatic injury, especially in spinal cord and brain.

Long Name

Activating Transcription Factor 6

Alternate Names

ACHM7, ATF6 alpha

Entrez Gene IDs

22926 (Human)

Gene Symbol

ATF6

UniProt

P18850

Additional ATF6 Products

Product Documents for ATF6 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for ATF6 Antibody - BSA Free

Manufactured by Genomic Antibody Technology™. GAT FAQs

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for ATF6 Antibody - BSA Free

Customer Reviews for ATF6 Antibody - BSA Free (1)

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  • ATF6 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Human primary fibroblast
    Species: Human
    Verified Customer | Posted 10/31/2020
    ATF6 in human primary fibroblasts. No band detected. Primary antibody incubation O/N 4ºC 1:1000 in BSA 3% PBST (0.1% Tween). Secondary ab: Donkey anti-rabit Alexa Fluor 647 1:5000 1h RT in BSA 3% PBST (0.1% Tween).
    TGX 12% gel.10 ug of protein loaded reduced with DTT. Primary antibody incubation O/N 4ºC 1:1000 in BSA 3% PBST (0.1% Tween). Secondary ab: Donkey anti-rabit Alexa Fluor 647 1:5000 1h RT in BSA 3% PBST (0.1% Tween).
    ATF6 Antibody - BSA Free NBP1-75478
    Bio-Techne Response
    Thank you for reviewing our product. We are sorry to that that this antibody did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction.

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Protocols

View specific protocols for ATF6 Antibody - BSA Free (NBP1-75478):

ATF6 Antibody:
Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for ATF6 Antibody - BSA Free

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    • Q: We are looking for an antibody specific to ATF6 alpha that will not cross-react with ATF6 beta. Would you please help confirm if any of the following are ATF6 alpha specific antibodies: NBP1-40256, NBP1-77251, NBP1-76675 & NBP1-41439?
      A: The antibodies NBP1-77251 and NBP1-76675 are specific to ATF6 alpha and should not cross-react with ATF6 beta.

    • Q: Will this antibody detect the cleaved portion of ATF6 (60-80kDa), as well as the non-cleaved portion (110 kDa)? We are using human lymphocyte cell lysates (samples of patients with leukemia) on a proteinsimple Wes machine.
      A: Thank you for contacting Novus regarding the ATF6 Antibody (NBP1-75478). We do see the full length in Simple Western but not the cleavage product. Nevertheless we assume you should see the 46kD cleavage product since the immunogen is located there but only under ER stress. The actual epitope is not mapped but the immunogen is with in the region of a.a. 200-250. Since the cleavage site is at 419-420 then we assume it is possible to detect this portion of the cleavge product but not the 27KD product without the immunogen sequence. 74 kD full length        aa1-420=46kD       aa420-670=27kD
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