Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR10881] to FOXA1 - BSA and Azide free
  • Suitable for: IHC-P, ICC/IF, Flow Cyt, WB, ChIP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-FOXA1 antibody [EPR10881] - BSA and Azide free
    See all FOXA1 primary antibodies

  • Description

    Rabbit monoclonal [EPR10881] to FOXA1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment within Human FOXA1 aa 350 to the C-terminus. The exact sequence is proprietary.
    Database link: P55317

  • Positive control

    • WB: Hap1, HeLa, SW480 and HepG2 whole cell lysate (ab7900). Mouse and rat lung lysates IHC-P: Human breast carcinoma & prostate tissue, mouse liver, Rat pancreas tissue. ICC/IF: PC-3 and HepG2 cells. ChIP: LNCaP (Human prostate carcinoma epithelial cell).

  • General notes

    Ab236011 is the carrier-free version of ab170933. 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

    ab236011 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Western blot - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    All lanes : Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : FOXA1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 52 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab170933).

      Lanes 1- 2: Merged signal (red and green). Green - ab170933 observed at 52 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab170933 was shown to react with FOXA1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261823 (knockout cell lysate ab256920) was used. Wild-type HeLa and FOXA1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab170933 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • ChIP - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    ChIP - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

    Chromatin was prepared from LNCaP cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab170933 (red), and 20µl of protein A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 5μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

  • Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

    Immunocytochemistry/ Immunofluorescence analysis of PC-3 (Human prostate adenocarcinoma epithelial cell) cells labeling FOXA1 with Purified ab170933 at 1:100 dilution (11 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor®594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor®488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat pancreas tissue sections labeling FOXA1 with Purified ab170933 at 1:1000 dilution (1.13 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylinwas used as a counterstain

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse liver tissue sections labeling FOXA1 with Purified ab170933 at 1:1000 dilution (1.13 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylinwas used as a counterstain

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling FOXA1 with Purified ab170933 at 1:1000 dilution (1.13 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylinwas used as a counterstain.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

  • Western blot - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Western blot - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    All lanes : Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : FOXA1 knockout HAP1 whole cell lysate
    Lane 3 : MCF7 whole cell lysate
    Lane 4 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 49 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab170933 observed at 52 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab170933 was shown to specifically react with FOXA1 in wild-type HAP1 cells as signal was lost in FOXA1 knockout cells. Wild-type and FOXA1 knockout samples were subjected to SDS-PAGE. Ab170933 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

  • Flow Cytometry - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Flow Cytometry - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

    Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling FOXA1 with purified ab170933 at 1/200 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling FOXA1 with unpurified ab170933 at 1/100 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

    Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

    Immunohistochemical analysis of paraffin-embedded Human prostate tissue labeling FOXA1 with unpurified ab170933 at 1/100 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

    Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

    Immunofluorescence analysis of HepG2 cells labeling FOXA1 with unpurified ab170933 at 1/100 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab170933).

  • Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)

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