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    products/primary-antibodies/foxa1-antibody-epr10881-chip-grade-ab170933.pdf

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Epigenetics and Nuclear Signaling Transcription Domain Families Forkhead Box FOXA
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

  • Datasheet
  • SDS
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Western blot - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • ChIP - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Western blot - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Western blot - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Flow Cytometry (Intracellular) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
  • Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

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 - ChIP Grade
  • Suitable for: ChIP, Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-FOXA1 antibody [EPR10881] - ChIP Grade
    See all FOXA1 primary antibodies
  • Description

    Rabbit monoclonal [EPR10881] to FOXA1 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ChIP, Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • WB: HeLa, Hap1, 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

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR10881
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Forkhead Box
    • FOXA
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

Associated products

  • Alternative Versions

    • Anti-FOXA1 antibody [EPR10881-14] (ab173287)
    • Alexa Fluor® 647 Anti-FOXA1 antibody [EPR10881] (ab196908)
    • Alexa Fluor® 488 Anti-FOXA1 antibody [EPR10881] (ab197235)
    • Anti-FOXA1 antibody [EPR10881] - BSA and Azide free (ab236011)
    • Anti-FOXA1 antibody [EPR10881-14] - BSA and Azide free (ab249749)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human FOXA1 knockout HeLa cell line (ab261823)
  • KO cell lysates

    • Human FOXA1 knockout HeLa cell lysate (ab256920)
  • Positive Controls

    • Hep G2 whole cell lysate (ab166833)
    • Mouse lung normal tissue lysate - total protein (ab29297)
    • SW480 whole cell lysate (ab3957)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab170933 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ChIP
Use 5 µg for 25 µg of chromatin.
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB (1)
1/1000 - 1/10000. Predicted molecular weight: 49 kDa.
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use 1/100 - 1/250

ICC/IF
1/100 - 1/250.
Notes
ChIP
Use 5 µg for 25 µg of chromatin.
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Predicted molecular weight: 49 kDa.
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use 1/100 - 1/250

ICC/IF
1/100 - 1/250.

Target

  • Function

    Transcription factor that is involved in embryonic development, establishment of tissue-specific gene expression and regulation of gene expression in differentiated tissues. Is thought to act as a 'pioneer' factor opening the compacted chromatin for other proteins through interactions with nucleosomal core histones and thereby replacing linker histones at target enhancer and/or promoter sites. Binds DNA with the consensus sequence 5'-[AC]A[AT]T[AG]TT[GT][AG][CT]T[CT]-3' (By similarity). Proposed to play a role in translating the epigenetic signatures into cell type-specific enhancer-driven transcriptional programs. Its differential recruitment to chromatin is dependent on distribution of histone H3 methylated at 'Lys-5' (H3K4me2) in estrogen-regulated genes. Involved in the development of multiple endoderm-derived organ systems such as liver, pancreas, lung and prostate; FOXA1 and FOXA2 seem to have at least in part redundant roles (By similarity). Modulates the transcriptional activity of nuclear hormone receptors. Is involved in ESR1-mediated transcription; required for ESR1 binding to the NKX2-1 promoter in breast cancer cells; binds to the RPRM promter and is required for the estrogen-induced repression of RPRM. Involved in regulation of apoptosis by inhibiting the expression of BCL2. Involved in cell cycle regulation by activating expression of CDKN1B, alone or in conjunction with BRCA1. Originally discribed as a transcription activator for a number of liver genes such as AFP, albumin, tyrosine aminotransferase, PEPCK, etc. Interacts with the cis-acting regulatory regions of these genes. Involved in glucose homeostasis.
  • Tissue specificity

    Highly expressed in prostate and ESR1-positive breast tumors. Overexpressed in esophageal and lung adenocarcinomas.
  • Sequence similarities

    Contains 1 fork-head DNA-binding domain.
  • Cellular localization

    Nucleus.
  • Target information above from: UniProt accession P55317 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3169 Human
    • Entrez Gene: 15375 Mouse
    • Entrez Gene: 25098 Rat
    • Omim: 602294 Human
    • SwissProt: P55317 Human
    • SwissProt: P35582 Mouse
    • SwissProt: P23512 Rat
    • Unigene: 163484 Human
    • Unigene: 4578 Mouse
    • Unigene: 10470 Rat
    see all
  • Alternative names

    • forkhead box A1 antibody
    • Forkhead box protein A1 antibody
    • FOX A1 antibody
    • FOXA1 antibody
    • FOXA1_HUMAN antibody
    • hepatocyte nuclear factor 3 alpha antibody
    • Hepatocyte nuclear factor 3-alpha antibody
    • HNF 3A antibody
    • HNF-3-alpha antibody
    • HNF-3A antibody
    • HNF3A antibody
    • MGC33105 antibody
    • TCF 3A antibody
    • TCF-3A antibody
    • TCF3A antibody
    • Transcription factor 3A antibody
    see all

Images

  • Western blot - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Western blot - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    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.

    Unpurified 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. 

  • ChIP - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    ChIP - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

    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).

  • Western blot - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Western blot - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    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?



    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.

  • Western blot - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Western blot - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    All lanes : Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933) at 1.1 µg/ml (purified)

    Lane 1 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse lung lysates
    Lane 3 : Rat lung lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 49 kDa
    Observed band size: 49 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling FOXA1 with unpurified ab170933 at 1/100 dilution. Heat mediated antigen retrieval was performed using citrate buffer pH 6.0 before commencing with IHC staining protocol.

  • Flow Cytometry (Intracellular) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Flow Cytometry (Intracellular) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

    Intracellular 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. 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

    Immunohistochemical analysis of paraffin-embedded Human prostate tissue labeling FOXA1 with unpurified ab170933 at 1/100 dilution. Heat mediated antigen retrieval was performed using citrate buffer pH 6.0 before commencing with IHC staining protocol.

  • Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)
    Anti-FOXA1 antibody [EPR10881] - ChIP Grade (ab170933)

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (21)

Publishing research using ab170933? Please let us know so that we can cite the reference in this datasheet.

ab170933 has been referenced in 21 publications.

  • Baca SC  et al. Reprogramming of the FOXA1 cistrome in treatment-emergent neuroendocrine prostate cancer. Nat Commun 12:1979 (2021). PubMed: 33785741
  • Golden EJ  et al. Onset of taste bud cell renewal starts at birth and coincides with a shift in SHH function. Elife 10:N/A (2021). PubMed: 34009125
  • Chen D  et al. PPM1G promotes the progression of hepatocellular carcinoma via phosphorylation regulation of alternative splicing protein SRSF3. Cell Death Dis 12:722 (2021). PubMed: 34290239
  • Centonze A  et al. Heterotypic cell-cell communication regulates glandular stem cell multipotency. Nature 584:608-613 (2020). PubMed: 32848220
  • Font A  et al. Immunohistochemistry-Based Taxonomical Classification of Bladder Cancer Predicts Response to Neoadjuvant Chemotherapy. Cancers (Basel) 12:N/A (2020). PubMed: 32635360
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

Western blot abreview for Anti-FOXA1 antibody [EPR10881]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T and Prostate cancer cells)
Gel Running Conditions
Reduced Denaturing (4-15% TGX BioRad)
Loading amount
200000 cells
Specification
HEK293T and Prostate cancer cells
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Jan 05 2016

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