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17-10258

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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Size/SKUAvailabilityPrice
25 assays
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$634.00

About This Item

UNSPSC Code:
12352203
NACRES:
NA.32
eCl@ss:
32160702
Clone:
monoclonal
Species reactivity:
human
Application:
ChIP, ICC, WB
Citations:
3

$634.00


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biological source

mouse

Quality Segment

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.
~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.

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This Item
171026717100611710237
description

from mouse

description

from rabbit, purified by affinity chromatography

description

from mouse

description

from mouse

clone

monoclonal

clone

polyclonal

clone

monoclonal

clone

monoclonal

species reactivity

human

species reactivity

mouse, human

species reactivity

human, Xenopus

species reactivity

rat, bovine, mouse, hamster, human

biological source

mouse

biological source

rabbit

biological source

mouse

biological source

mouse

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

UniProt accession no.

Q9Y261

UniProt accession no.

P55317

UniProt accession no.

Q01094

UniProt accession no.

Q92769

technique(s)

ChIP: suitable, western blot: suitable, immunocytochemistry: suitable

technique(s)

ChIP: suitable, immunocytochemistry: suitable, western blot: suitable

technique(s)

ChIP: suitable (ChIP-seq), immunoprecipitation (IP): suitable

technique(s)

ChIP: suitable, western blot: suitable


Storage Class

10 - Combustible liquids

wgk

WGK 1



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Questions

  1. Could you confirm whether this antibody is free from cross-reactivity with FOXA1 for upcoming ChIP and CRISPR/Cas9 experiments?

    1 answer
    1. The proteins belonging to the Foxa family (Foxa1, Foxa2, and Foxa3) share very high sequence homology within the DNA binding domain. Literature indicates that FOXA2 interacts with both FOXA1 and FOXA3 in vivo, while FOXA1 and FOXA3 do not appear to interact. The immunogen for this antibody is a recombinant protein corresponding to the full length of human FOXA2. The epitope this antibody binds to has not been mapped. 17-10258 Sigma-Aldrich ChIPAb+™ FOXA2 is a ChIP application validated antibody. However, no specific assays have been performed to determine or rule out any possibility of this antibody cross-reacting with FOXA1. We apologize for any inconvenience this may cause.

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