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

产品说明

一般描述

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.

特异性

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

免疫原

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

应用

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

质量

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.

目标描述

~55 kDa

外形

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
Protein G Purified
Format: Purified

分析说明

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

法律信息

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

免责声明

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

基本信息

产品性质

安全信息