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ChIPAb+ E2F-3 - ChIP Validated Antibody and Primer Set
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Species Reactivity Key Applications Host Format Antibody Type
H, M, R WB, ChIP Mouse Purified Monoclonal Antibody
Description:
ChIPAb+ E2F-3 - 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+ E2F-3 set includes the E2F-3 antibody, a negative control normal mouse IgG, and qPCR primers which amplify a 100 bp region of human CDC2 promoter. The E2F-3 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of E2F-3-associated chromatin.
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Specificity:
E2F-3
Molecular Weight:
~58 kDa
Epitope:
Unknown
Immunogen:
Recombinant full-length human E2F-3 protein
Isotype:
IgG2a
Background Information:
E2F transcription factors are functionally regulated by binding to Rb p110, p107, and p130. E2F 3 is regulated by complex formation with Rb p110. E2F family members bind DNA as heterodimers with members of the DP family of polypeptides.
Species Reactivity:
Human
Mouse
Rat
Species Reactivity Note:
Human and mouse. Weak reactivity with rat.
Application Notes:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (4 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Mouse IgG, or 1 µg Anti-E2F-3 and the Magna ChIP™ A Kit (Cat. # 17-610). Successful immunoprecipitation of E2F-3 associated DNA fragments was verified by qPCR using ChIP Primers, human CDC2 promoter as a positive locus, and a intergenic region near human CALML5 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 Blot Analysis:
Representative lot data.
Lysates from HeLa cells probed with anti-E2F-3 at 2 μg/mL.
Arrow indicates E2F-3 (~58 kDa) (Figure 3).
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Control:
Includes normal mouse IgG and primers specific for human CDC2 coding region.
Quality Assurance:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (4 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Mouse IgG, or 1 µg Anti-E2F-3 and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of E2F-3 associated DNA fragments was verified by qPCR using ChIP Primers, human CDC2 promoter (Figure 1).
Please refer to the EZ-Magna ChIP™ A (Cat. # 17-408) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details.
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Purification Method:
Protein A purfied
Presentation:
Anti-E2F-3 (mouse monoclonal), . One vial containing 25 ug of protein A purified mouse IgG2a in ~18 |