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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H, M | WB, ICC, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-10267 |
| Trade Name |
|
| Description | ChIPAb+™ FOXA1 Antibody |
| 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+™ 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. |
| Alternate Names |
|
| Background Information | HNF-3A, Hepatocyte nuclear factor 3-alpha (also called FOXA1) is member of the Forkhead family of winged-helix transcription factors. This protein family has around 50 known transcription factors and play an important role in the regulation of cell differentiation, organogenesis, and gene expression. High levels of expression of HNF-3A has been observed in tumors arising from both the prostate and breast. Here it is believed this protein interacts with the androgen receptor in prostate tumors and the estrogen receptor α in breast tumors. In the case of ERα, HNF-3A interacts with cis-regulatory regions in heterochromatin and enhances the interaction of ERα with chromatin. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Control |
|
| Presentation | Anti-FOXA1 (Rabbit Polyclonal). One vial containing 50 µL of purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH 7.4, 150 mM NaCl) with 0.05% sodium azide before the addition of glycerol to 30%. Store at -20°C. Concentration: 0.35 mg/mL Normal Rabbit IgG. One vial containing 125 µg Rabbit IgG in 125 µL storage buffer containing 0.05% 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 |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ FOXA1 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
|
| Application Notes | 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) |
| Biological Information | |
|---|---|
| Immunogen | Recombinant protein corresponding to the entire protein of human FOXA1 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Rabbit |
| Specificity | This antibody recognizes HNF-3A/FOXA1. |
| Species Reactivity |
|
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
|
| Entrez Gene Summary | This gene encodes a member of the forkhead class of DNA-binding proteins. These hepatocyte nuclear factors are transcriptional activators for liver-specific transcripts 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. [provided by RefSeq]. |
| Gene Symbol |
|
| Purification Method | Affinity Purfied |
| UniProt Number |
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| UniProt Summary | FUNCTION: 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. SUBCELLULAR LOCATION: Nucleus. SEQUENCE SIMILARITIES: Contains 1 fork-head DNA-binding domain. |
| Molecular Weight | ~51 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | 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 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 (Figure 1). Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. |
| Usage Statement |
|
| 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: 2 μL of antibody per chromatin immunoprecipitation (dependent upon biological context). |