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ChIPAb+ E2F-1 - ChIP Validated Antibody and Primer Set
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Species Reactivity Key Applications Host Format Antibody Type
H, Xn IP, ChIP-seq, ChIP Mouse Purified Monoclonal Antibody
Description:
ChIPAb+ E2F-1 - 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-1 set includes the E2F-1 antibody, a negative control normal mouse IgG, and qPCR primers which amplify a 100 bp region of human CDC2 promoter. The E2F-1 and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of E2F-1-associated chromatin.
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Specificity:
The epitopes have been mapped to amino acids 1-89 and to amino acids 342-386.
Molecular Weight:
~55 kDa
Epitope:
a.a. 1-89 & 342-386
Immunogen:
GST fusion protein corresponding to full-length human E2F-1.
Isotype:
IgG
Background Information:
E2F-1 was identified as a cellular protein with DNA binding activity associated with the adenovirus E2 gene promoter. The original member of the E2F family of transcription factors, now consisting of five, was identified through its association with the retinoblastoma protein, which controls its activity. E2F-1 is cell cycle regulated with very low levels in early G1 then increasing levels as cells move from G1 to S with highest levels of protein at the G1/S boundary, which is consistent with its role in S-phase entry.
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Species Reactivity:
Human
Xenopus
Species Reactivity Note:
Human and xenopus.
Application Notes:
Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µL of either Normal Mouse IgG or 1 µL of Anti-E2F-1 and the Magna ChIP™ G Kit (Cat. # 17-611).
Successful immunoprecipitation of E2F-1-associated DNA fragments was verified by qPCR using ChIP Primers, human CDC2 promoter as a positive locus, and an intergenic region near human CALML5 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.
HeLa nuclear extract was resolved by electrophoresis, transferred to PVDF and probed with Anti-E2F-1 (1.0 µg/mL).
Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection
system.
Arrow indicates E2F-1 (~55 kDa) (Please see figures).
Immunoprecipitation: 4 μg of a previous lot of this antibody immunoprecipitated E2F-1 from HeLa nuclear extract.
Electrophoretic Mobility Shift Assay (EMSA): A previous lot of the antibody has been shown to gel shift the E2F-1/DP-1/DNA complex.
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