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ChIPAb+ TCF-4 - ChIP Validated Antibody and Primer Set
Species Reactivity Key Applications Host Format Antibody Type
H, M WB, DB, ChIP Mouse Purified Monoclonal Antibody
Description:
ChIPAb+ TCF-4 - ChIP Validated Antibody and Primer Set
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Trade Name:
Upstate (Millipore)
Product 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+ TCF-4 set includes the TCF-4 antibody, a negative control normal mouse IgG, and qPCR primers which amplify a 198 bp region of human SP5 promoter. The TCF-4 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of TCF-4-associated chromatin.
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Specificity:
This antibody recognizes TCF-4
Molecular Weight:
~66 kDa
Immunogen:
Amino acids 31-331 of human TCF-4.
Isotype:
IgG2a
Background Information:
TCF4, also known as TCF7L2, is expressed widely during development. Gene targeting study indicates that it is required to maintain the crypt stem cells of the small intestine. TCF4 has many different splicing isoforms and they are expressed differentially in tissues and in cancers of different stages. Studies also indicate that variant of the TCF4 gene confers an increased risk of type 2 diabetes.
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Species Reactivity:
Human
Mouse
Species Reactivity Note:
Human and mouse
Application Notes:
Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HT29 cells (3 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of Normal Mouse IgG or 2 µg Anti-TCF-4 and the Magna ChIP™ G Kit (Cat. # 17-611).
Successful immunoprecipitation of TCF-4 associated DNA fragments was verified by qPCR using ChIP Primers, SP5 Promotor as a positive locus, and GAPDH promoter primers as a negative locus (Please see figures). 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™ G (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
Western Blot Analysis:
Representative lot data.
Cos-1 cells were transfected with TCF-4 cDNA in pUSE (lane 2) or pUSE (lane 1). RIPA lysates were prepared, resolved by electrophoresis, transferred to nitrocellulose and probed with anti-TCF-4 (1 mg/mL).
Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates TCF-4 (~66 kDa) (Please see figures).
Immunocytochemistry/Immunohistochemistry:
A previous lot of this antibody is reported to show positive immunostaining for TCF-4 in cells fixed with 4% paraformaldehyde/PBS (Amir, A. L., 2003).
Immunoprecipitation:
A previous lot of this antibody was reported to immunoprecipitate TCF-4 from SW620 whole cell lysate as a complex with Beta-Catenin (Persad, S., 2001).
Supershift Assay (EMSA):
A previous lot of this antibody was reported to supershift TCF-4/DNA complexes in electrophoretic mobility shift assays (EMSA) (Amir, A. L., 2003).< |