Invitrogen

FOXA1 Polyclonal Antibody

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated. View Details

6 Published Figures

11 References

View all (25) FOXA1 antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

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FOXA1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of FOXA1 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with FOXA1 Polyclonal Antibody (Product # PA5-27157) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

FOXA1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of FOXA1 in the nucleus of paraffin-embedded human cervical carcinoma using a FOXA1 polyclonal antibody (Product # PA5-27157) at a dilution of 1:500.

FOXA1 Antibody in Western Blot (WB) — Western blot was performed using Anti-FOXA1 Polyclonal Antibody(Product # PA5-27157) and a 51kDa band corresponding to FOXA1 was observed across cell lines tested except SW480, HCT 116 and SH-SY5Y which is reported to be low. Whole Cell Extract (1% SDS) (30 µg lysate) of MCF7 (Lane 1), T-47D (Lane 2), HT-29 (Lane 3), SW480 (Lane 4), HCT 116 (Lane 5) and SH-SY5Y (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:3000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

FOXA1 Antibody in Immunoprecipitation (IP) — PA5-27157 immunoprecipitates FOXA1 protein in IP experiments. IP Sample: HepG2 whole cell lysate/extract A: 40 µg whole cell lysate/extract of FOXA1 protein expressing HepG2 cells B: Control with 2.5 µg of pre-immune rabbit IgG C: Immunoprecipitation of FOXA1 by 2.5 µg of FOXA1 antibody (Product # PA5-27157) on 10% SDS-PAGE.

FOXA1 Antibody in ChIP assay (ChIP) — Chromatin Immunoprecipitation (ChIP) assay of endogenous FOXA1 protein using FOXA1 Antibody: ChIP was performed using Anti-FOXA1 Polyclonal Antibody (Product # PA5-27157, 2.5 µg) on sheared chromatin from LnCaP cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to KLK3, SLµg, TMPRSS2 (Active) and SAT2 satellite repeats (Inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

FOXA1 Antibody — Figure 5 FOXA1 is recruited by the AR to the enhancers and regulates the transcription of TMPRSS2 and PRCAT38 . ( a , b ) ChIP-qPCR before and after DHT treatment showing FOXA1 enrichment over the enhancer regions in LNCaP and VCaP. ( c ) Western blot showing the protein level of FOXA1 and H3K27ac after FOXA1 knockdown with or without DHT treatment. ( d - e ) qPCR showing the down-regulation of FOXA1 , TMPRSS2 and PRCAT38 after FOXA1 knockdown with or without DHT treatment. Data are shown as mean +- SD (n = 3). {KD}

FOXA1 Antibody in Western Blot, Immunoprecipitation (WB, IP) — Extended Data Figure 4 Functional impact of FOXA1 mutations on oncogenic AR-signaling. a) Immunoblot showing expression of endogenous and V5-tagged exogenous FOXA1 proteins in dox-inducible 22RV1 cells transfected with distinct UTR-specific FOXA1 siRNAs (#3-5) or a non-targeting control siRNA (siNC). These results represent 2 independent experiments. Incucyte growth curves of 22RV1 cells overexpressing empty vector (control), WT, or mutant FOXA1 variants upon treatment with UTR-specific FOXA1-targeting siRNAs (n = 5 biological replicates). Mean +- s.e.m are shown. b) Immunoblots confirming stable overexpression of the WT AR protein in HEK293 and PC3 cells. c, d) Co-immunoprecipitation assay of indicated recombinant FOXA1 variants using a V5-tag antibody in (c) HEK293-AR and (d) PC3-AR cells. eGFP is a negative control; FL is the full-length WT FOXA1. d168 and d358 are truncated FOXA1 variants with only the first 168aa (i.e. before the Forkhead domain) or 358aa of FOXA1 protein. H247Q and R261G are missense class1 mutant variants. e) Immunoblots confirming comparable expression of AR and recombinant FOXA1 variants in AR reporter assay-matched HEK293 lysates. Immunoblots show representative results from 2-3 independent experiments and class1 and class2 mutants serve as biological replicates. For all gel source data ( a,b-e ), see Supplementary Figure 1 . f) AR dual-luciferase reporter assays with transient overexpression of indicated FOXA1 variant in HEK293-AR cells with or wit

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:3,000

View 2 publications 2 publications

Immunohistochemistry (IHC)

View 3 publications 3 publications

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:1,000

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

1:100-1:1,000

Immunoprecipitation (IP)

1:100-1:500

View 1 publication 1 publication

ChIP assay (ChIP)

2.5 µg/10^6 cells

View 3 publications 3 publications

Dot blot (DB)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Human

Published species

Human

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide corresponding to a region within amino acids 412 and 472 of FOXA1

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7, with 20% glycerol

Contains

0.01% thimerosal

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2544633

Product Specific Information

Recommended positive controls: A431, HepG2.

Predicted reactivity: Mouse (100%), Rat (100%), Zebrafish (100%), Xenopus laevis (100%), Bovine (100%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

This gene encodes a member of the forkhead class of DNA-binding proteins. These hepatocyte nuclear factors are transcriptional activators for liver-specific transcripts such as albumin and transthyretin, and they also interact with chromatin. Similar family members in mice have roles in the regulation of metabolism and in the differentiation of the pancreas and liver.

⚠WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause birth defects or other reproductive harm. For more information go to www.P65Warnings.ca.gov.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Ci-fkh; fkd7; fkh; Forkhead box protein A1; forkhead-7; FoxA4a; Hepatocyte nuclear factor 3-alpha; HNF-3-alpha; HNF-3A; HNF3-alpha; HNF3alpha; MGC33105; MGC52590; MGC75967; pintallavis; TCF-3A; Transcription factor 3A; xfkh1; xfkh2

Gene Aliases: FOXA1; HNF3A; TCF3A

UniProt ID: (Human) P55317

Entrez Gene ID: (Human) 3169

Molecular Function: winged helix/forkhead transcription factor