Description
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+ FOXA2 set includes the FOXA2 antibody, a Normal Mouse IgG, and control primers which amplify a 138 bp region of ChIP Primers, Mouse Hnf4α enhancer. The FOXA2 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of FOXA2-associated chromatin.
Also known as hepatocyle nuclear factor 3-beta (HNF-3B), Forkhead box protein A2 (FOXA2) belongs to the forkhead class of DNA-binding proteins. This family of transcription factors influences gene expression in endodermally-derived tissues such as lung, liver, stomach, pancreas, and intestine. Foxa2 has also been detected in ectodermally-derived tissues such as kidney, uterus and vagina, and seminal and coagulating glands during development. Several studies show that FOXA proteins help facilitiate the binding of several nuclear receptors to their respective targets in a context-dependent manner, and are thereby implicated in various functions including cellular maintenance, differentiation, metabolism, and organogenesis. Aberrant function of FOXA proteins are attributed to diseases states such as diabetes and Parkinson′s disease.
~50 kDa observed
Immunogen
Recombinant protein corresponding to the human FOXA2.
Application
Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from Mouse liver tissues (5 mg tissue equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µL of either Normal Mouse IgG, or 4 µL of Anti-FOXA2 and the Magna ChIP G Kit (Cat. # 17-611). Successful immunoprecipitation of FOXA2 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. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
HepG2 cell lysate was probed with Anti-FOXA2 (1:500 dilution). Proteins were visualized using a Goat Anti-Mouse IgG secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates FOXA2 (~50 kDa). (Figure 3).
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors
This ChIPAb+ FOXA2 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Packaging
25 assays per set. Recommended use: 4 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
Physical form
Anti-FOXA2 (mouse monoclonal),. One vial containing 100 µL of purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4) and 150 mM NaCl with 0.05% sodium azide before the addition of glycerol to 30%. Store at -20C.
Concentration: 0.175 mg/mL
Normal Mouse IgG. One vial containing 125 µg of purified mouse IgG in 125 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.
ChIP Primers, Mouse Hnf4α enhancer. One vial containing 75 μL of 5 μM of each primer specific for Mouse Hnf4α enhancer. Store at -20°C.
FOR: TTC CAG CTG CCT TTA TCT CCC TGT
REV: TCT CCA CAC ATG TCC AGC AGC CT
Format: Purified
Protein G
Preparation Note
Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
Analysis Note
Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from Mouse liver tissues (5 mg tissue equivalents per IP) were subjected to chromatin immunoprecipitation using 4 µL of either Normal Mouse IgG,or 4 µL of Anti-FOXA2 and the Magna ChIP® G Kit (Cat. # 17-611). Successful immunoprecipitation of FOXA2 associated DNA fragments was verified by qPCR using ChIP Primers, Mouse Hnf4α enhancer (Figure 1).
Please refer to the EZ-Magna ChIP G (Cat. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Control
Includes normal mouse IgG and primers specific for Mouse Hnf4α enhancer.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
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.
