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17-10258
Sigma-AldrichChIPAb+™ FOXA2 - ChIP Validated Antibody and Primer Set
This ChIPAb+ FOXA2 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
More>>This ChIPAb+ FOXA2 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. Less<<
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ChIPAb+™ FOXA2 - ChIP Validated Antibody and Primer Set
Overview
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.
Alternate Names
Hepatocyte nuclear factor 3-beta
HNF-3-beta
HNF-3B
Forkhead box protein A2
Transcription factor 3B
TCF-3B
Background Information
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.
References
Product Information
Format
Purified
Control
Includes normal mouse IgG and primers specific for Mouse Hnf4α enhancer.
Presentation
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
This ChIPAb+ FOXA2 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Key Applications
Western Blotting
Immunocytochemistry
Chromatin Immunoprecipitation (ChIP)
Application Notes
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).
Biological Information
Immunogen
Recombinant protein corresponding to the human FOXA2.
Concentration
Please refer to the Certificate of Analysis for the lot-specific concentration.
Host
Mouse
Isotype
IgG1κ
Species Reactivity
Human
Mouse
Species Reactivity Note
Demonstrated to react with human. Predicted to react with mouse based on 100% sequence homology.
This gene encodes a member of the forkhead class of DNA-binding proteins. These hepatocyte nuclear factors are transcriptional activators for liver-specific genes 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. This gene has been linked to sporadic cases of maturity-onset diabetes of the young. Transcript variants encoding different isoforms have been identified for this gene. [provided by RefSeq].
FUNCTION: Transcription factor that is involved in embryonic development, establishment of tissue-specific gene expression and regulation of gene expression in differentiated tissues. Is thought to act as a 'pioneer' factor opening the compacted chromatin for other proteins through interactions with nucleosomal core histones and thereby replacing linker histones at target enhancer and/or promoter sites. Binds DNA with the consensus sequence 5'-[AC]A[AT]T[AG]TT[GT][AG][CT]T[CT]-3' (By similarity). In embryonic development is required for notochord formation. Involved in the development of multiple endoderm-derived organ systems such as the liver, pancreas and lungs; FOXA1 and FOXA2 seem to have at least in part redundant roles. Originally discribed as a transcription activator for a number of liver genes such as AFP, albumin, tyrosine aminotransferase, PEPCK, etc. Interacts with the cis-acting regulatory regions of these genes. Involved in glucose homeostasis; regulates the expression of genes important for glucose sensing in pancreatic beta-cells and glucose homeostasis. Involved in regulation of fat metabolism. Binds to fibrinogen beta promoter and is involved in IL6-induced fibrinogen beta transcriptional activation.
SUBUNIT STRUCTURE: Binds DNA as a monomer. Binds TLE1 (By similarity). Interacts with FOXA1 and FOXA3. Interacts with PRKDC.
SUBCELLULAR LOCATION: Nucleus. Cytoplasm. Note: Shuttles between the nucleus and cytoplasm in a CRM1-dependent manner; in response to insulin signaling via AKT1 is exported from the nucleus.
PTM: Phosphorylation on Thr-156 abolishes binding to target promoters and subsequent transcription activation upon insulin stimulation (By similarity).
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.
Usage Statement
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 and Shipping Information
Storage Conditions
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.
Packaging Information
Material Size
25 assays
Material Package
25 assays per set. Recommended use: 4 μL of antibody per chromatin immunoprecipitation (dependent upon biological context).
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
17-10258
04053252756542
Documentation
ChIPAb+™ FOXA2 - ChIP Validated Antibody and Primer Set SDB
Global analysis of in vivo Foxa2-binding sites in mouse adult liver using massively parallel sequencing. Wederell, Elizabeth D, et al. Nucleic Acids Res., 36: 4549-64 (2008)
2008
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 profile of in vivo Foxa2-binding sites in the adult liver. More than 65% of the approximately 11.5 k genomic sites associated with Foxa2 binding, mapped to extended gene regions of annotated genes, while more than 30% of intragenic sites were located within first introns. 20.5% of all sites were further than 50 kb from any annotated gene, suggesting an association with novel gene regions. QPCR analysis demonstrated a strong positive correlation between peak height and fold enrichment for Foxa2-binding sites. We measured the relationship between Foxa2 and liver gene expression by overlapping Foxa2-binding sites with a SAGE transcriptome profile, and found that 43.5% of genes expressed in the liver were also associated with Foxa2 binding. We also identified potential Foxa2-interacting transcription factors whose motifs were enriched near Foxa2-binding sites. Our comprehensive results for in vivo Foxa2-binding sites in the mouse liver will contribute to resolving transcriptional regulatory networks that are important for adult liver function.
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