Analysis Note
ControlIncludes normal rabbit IgG and primers specific for Mouse Hnf4α enhancer.
Application
Research Sub CategoryTranscription Factors
Research CategoryEpigenetics & Nuclear Function
Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from Mouse liver tissue (1 mg tissue equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µl of either normal rabbit IgG, or 2 µl Anti-FOXA1 (Part No.CS207368) and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of FoxA1 associated DNA fragments was verified by qPCR using ChIP Primers, Mouse Hnf4α enhancer as a positive locus, and mouse Hnf4α promoter as a negative locus. (Figure 2). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blotting Analysis:
Representative lot data.
Human pancreas tissue lysate was resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-FOXA1 (0.5 µg/mL).
Proteins were visualized using a Donkey Anti-Rabbit IgG conjugated to HRP and chemiluminescence detection system. (Figure 3)
Arrow indicates FOXA1 (~51 kDa).
Immunocytochemistry Analysis:
Representative lot data.
Confocal fluorescent analysis of NIH/3T3 and HeLa cells using Anti-HNF-3A/FOXA1 (Red). Actin filaments have been labeled with Alexa Fluor™ 488 Dye-Phalloidin (Green). Nucleus is stained with DAPI (Blue). This antibody positively stains nucleus. (Figure 4)
This ChIPAb+ FOXA1 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
General description
All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+™ FOXA1 set includes the FOXA1 antibody, a Normal Rabbit IgG, and control primers which amplify a 138 bp region of ChIP Primers, Mouse Hnf4α enhancer. The FOXA1 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of FOXA1-associated chromatin.
HNF-3A, Hepatocyte nuclear factor 3-alpha (also called FOXA1) is member of the Forkhead family of winged-helix transcription factors