Recombinant
RabMAb

Recombinant Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22919-71] to FOXA2 - ChIP Grade – BSA and Azide free
  • Suitable for: ICC/IF, Flow Cyt, ChIP, IP, IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free
    See all FOXA2 primary antibodies

  • Description

    Rabbit monoclonal [EPR22919-71] to FOXA2 - ChIP Grade – BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody is suitable for mouse and rat for IHC but not other applications.

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, ChIP, IP, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human FOXA2 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9Y261

  • Positive control

    • WB: Human colon cancer, PC-3 and SW480 lysates. IHC-P: Human prostate neuroendocrine carcinoma, Mouse liver, Rat colon and mouse testis tissues. ICC/IF: PC-3 cells. Flow Cyt: PC-3 cells. IP: PC-3 whole cell lysate.

  • General notes

    Ab259272 is the carrier-free version of ab256493. 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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab259272 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab259272 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
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB Use at an assay dependent concentration.

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). In embryonic development is required for notochord formation. Involved in the development of multiple endoderm-derived organ systems such as the liver, pancreas and lungs; FOXA1 and FOXA2 seem to have at least in part redundant roles. 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; regulates the expression of genes important for glucose sensing in pancreatic beta-cells and glucose homeostasis. Involved in regulation of fat metabolism. Binds to fibrinogen beta promoter and is involved in IL6-induced fibrinogen beta transcriptional activation.

  • Sequence similarities

    Contains 1 fork-head DNA-binding domain.

  • Post-translational
    modifications

    Phosphorylation on Thr-156 abolishes binding to target promoters and subsequent transcription activation upon insulin stimulation.

  • Cellular localization

    Nucleus. Cytoplasm. Shuttles between the nucleus and cytoplasm in a CRM1-dependent manner and in response to insulin signaling via AKT1 is exported from the nucleus.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Forkhead box A2 antibody
    • Forkhead box protein A2 antibody
    • FOX A2 antibody
    • foxa2 antibody
    • FOXA2_HUMAN antibody
    • Hepatic nuclear factor 3 beta antibody
    • Hepatocyte nuclear factor 3 antibody
    • Hepatocyte nuclear factor 3 beta antibody
    • Hepatocyte nuclear factor 3-beta antibody
    • HNF 3B antibody
    • HNF-3-beta antibody
    • HNF-3B antibody
    • HNF3B antibody
    • MGC19807 antibody
    • TCF 3B antibody
    • TCF-3B antibody
    • TCF3B antibody
    • Transcription factor 3B antibody
    see all

Images

  • Immunoprecipitation - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)
    Immunoprecipitation - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)

    FOXA2 was immunoprecipitated from 0.35 mg PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate 10ug with ab256493 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256493 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate 10ug

    Lane 2: ab256493 IP in PC-3 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab256493 in PC-3 whole cell lysate

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

    Exposure time: 15 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256493).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling FOXA2 with ab256493 at 1/1000 dilution (0.58ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Negative control: No staining in mouse testis (PMID:15567715). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256493).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)

    Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling FOXA2 with ab256493 at 1/1000 dilution (0.58ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in rat colon (PMID:15567715). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256493).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling FOXA2 with ab256493 at 1/1000 dilution (0.58ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in mouse liver (PMID: 15567715). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256493).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)

    Immunohistochemical analysis of paraffin-embedded Human prostate neuroendocrine carcinoma tissue labeling FOXA2 with ab256493 at 1/1000 dilution (0.58ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human prostate neuroendocrine carcinoma (PMID:18156212;28621319). Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256493).

  • Immunocytochemistry/ Immunofluorescence - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)
    Immunocytochemistry/ Immunofluorescence - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-3 (human prostate adenocarcinoma epithelial cell) cells labelling FOXA2 with ab256493 at 1/100 (5.8 ug/ml) dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear staining in PC-3 cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Ab256493 anti- FOXA2 ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 ug/ml) dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256493).

  • Flow Cytometry - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)
    Flow Cytometry - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized LNCaP (Human prostate carcinoma epithelial cell, Left) / PC-3 (Human prostate adenocarcinoma epithelial cell, Right) cells labelling FOXA2 with ab256493 at 1/600 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    Negative control: LNCaP (28621319,16001449).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab256493).

  • ChIP - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)
    ChIP - Anti-FOXA2 antibody [EPR22919-71] - ChIP Grade – BSA and Azide free (ab259272)

    Chromatin was prepared from HepG2 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min. 
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab256493 (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: PMC6515469.

    *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 (ab256493).

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab259272? Please let us know so that we can cite the reference in this datasheet.

ab259272 has not yet been referenced specifically in any publications.

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