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Key Documents

17-10061

Sigma-Aldrich

ChIPAb+ E2F-1 - ChIP Validated Antibody and Primer Set

from mouse

Sinônimo(s):

Transcription factor E2F1, Retinoblastoma-binding protein 3, PRB-binding protein E2F-1

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About This Item

Código UNSPSC:
12352203
eCl@ss:
32160702
NACRES:
NA.52

fonte biológica

mouse

Nível de qualidade

clone

monoclonal

reatividade de espécies

human, Xenopus

fabricante/nome comercial

ChIPAb+
Upstate®

técnica(s)

ChIP: suitable (ChIP-seq)
immunoprecipitation (IP): suitable

Isotipo

IgG

nº de adesão NCBI

nº de adesão UniProt

Condições de expedição

dry ice

Descrição geral

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.

Especificidade

The epitopes have been mapped to amino acids 1-89 and to amino acids 342-386.

Imunogênio

Epitope: a.a. 1-89 & 342-386

Aplicação

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.
This ChIPAb+ E2F-1 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Qualidade

Chromatin Immunoprecipitation:
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 (Please see figures).

Please refer to the EZ-Magna ChIP G (Cat. # 17-409) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Descrição-alvo

~55 kDa

forma física

Anti-E2F-1 (mouse monoclonal). One vial containing 25 µg of protein G purified monoclonal mixed IgG in 25 µL of buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl with 0.05% sodium azide. Frozen solution. Store at -20°C.

Nornal Mouse IgG. One vial containing 25 µg of purified IgG in 25 µL of storage buffer containing 0.1% sodium azide. Store at -20°C.

ChIP Primers, CDC2 promoter. One vial containing 75 μL of 5 μM each primer specific for human CDC2 promoter. Store at -20°C.
FOR: CGC CCT TTC CTC TTT CTT TC
REV: ATC GGG TAG CCC GTA GAC TT
Format: Purified

Nota de análise

Control
Includes negative control normal mouse IgG and primers specific for human CDC2 promoter.

Informações legais

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Código de classe de armazenamento

10 - Combustible liquids


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