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17-10258

Sigma-Aldrich

ChIPAb+ FOXA2 - ChIP Validated Antibody and Primer Set

from mouse

Synonym(s):

Hepatocyte nuclear factor 3-beta, HNF-3-beta, HNF-3B, Forkhead box protein A2, Transcription factor 3B, TCF-3B

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32

biological source

mouse

Quality Level

clone

monoclonal

species reactivity

human

species reactivity (predicted by homology)

mouse (based on 100% sequence homology)

manufacturer/tradename

ChIPAb+
Upstate®

technique(s)

ChIP: suitable
immunocytochemistry: suitable
western blot: suitable

isotype

IgG1κ

NCBI accession no.

UniProt accession no.

shipped in

dry ice

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.

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).

Quality

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.

Target description

~50 kDa observed

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

Storage and Stability

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

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.

Storage Class Code

10 - Combustible liquids


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Elizabeth D Wederell et al.
Nucleic acids research, 36(14), 4549-4564 (2008-07-10)
Foxa2 (HNF3 beta) is a one of three, closely related transcription factors that are critical to the development and function of the mouse liver. We have used chromatin immunoprecipitation and massively parallel Illumina 1G sequencing (ChIP-Seq) to create a genome-wide
Shuai Gao et al.
Nature genetics, 52(10), 1011-1017 (2020-09-02)
FOXA1 functions as a pioneer transcription factor by facilitating the access to chromatin for steroid hormone receptors, such as androgen receptor and estrogen receptor1-4, but mechanisms regulating its binding to chromatin remain elusive. LSD1 (KDM1A) acts as a transcriptional repressor
Ingo Burtscher et al.
International journal of molecular sciences, 22(7) (2021-04-04)
Nkx6-1 is a member of the Nkx family of homeodomain transcription factors (TFs) that regulates motor neuron development, neuron specification and pancreatic endocrine and β-cell differentiation. To facilitate the isolation and tracking of Nkx6-1-expressing cells, we have generated a novel

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