Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22690-84] to ATF6 - BSA and Azide free
  • Suitable for: WB, IHC-P, ChIP, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free
    See all ATF6 primary antibodies

  • Description

    Rabbit monoclonal [EPR22690-84] to ATF6 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ChIP, IPmore details
    Unsuitable for: Flow Cyt or ICC/IF

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1 and HeLa whole cell lysates. IHC-P: Human kidney tissue. IP: HeLa whole cell lysate. ChIP: Chromatin prepared from RAW 264.7 (treated with tunicamycin) and HeLa (treated with thapsigargin) cells.

  • General notes

    ab263955 is the carrier-free version of ab227830. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab263955 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)
    Western blot - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)
    All lanes : Anti-ATF6 antibody [EPR22690-84] - ChIP Grade (ab227830) at 1/1000 dilution

    Lane 1 : Wild type HAP1 whole cell lysate
    Lane 2 : F6 knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 74 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?



    ab227830 was shown to specifically react with ATF6 in wild-type HAP1 cells as signal was lost in ATF6 knockout cells. Wild-type and ATF6 knockout samples were subjected to SDS-PAGE. ab227830 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/20,000 dilution for 1 hour at room temperature before imaging.
     
    Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
     
    Exposure time 62 seconds.
     
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227830).

     

  • Immunoprecipitation - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)
    Immunoprecipitation - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)

    ATF6 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab227830 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab227830 1/1000 dilution (0.5 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg

    Lane 2: ab254324 IP in HeLa whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227830 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 min

    Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227830).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)

    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling ATF6 with ab227830 at 1/1000 dilution (0.566 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human kidney (PMID: 25725420) is observed. The section was incubated with ab227830 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227830).

     

  • ChIP - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)
    ChIP - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)

    Chromatin was prepared from HeLa cells treated with thapsigargin (1 uM, 1 h) according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30 mins and then formaldehyde for 10 min. 
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab227830 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). 
    Primers and probes are from paper PMID: 17535801


    *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227830).

  • ChIP - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)
    ChIP - Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)

    Chromatin was prepared from RAW 264.7 cells treated with tunicamycin (5 ug/ml, 4 h) according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30 mins and then formaldehyde for 10 min. 
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab227830 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). 
    Primers and probes are from paper PMCID: PMC5179193.


    *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227830).

  • Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)
    Anti-ATF6 antibody [EPR22690-84] - BSA and Azide free (ab263955)

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