17-10061

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

from mouse

• 別名: Transcription factor E2F1, Retinoblastoma-binding protein 3, PRB-binding protein E2F-1
• UNSPSCコード: 12352203
• eCl@ss: 32160702
• NACRES: NA.52

特性

• 由来生物: mouse
• 品質水準: 100
• クローン: monoclonal
• 交差性: human, Xenopus
• メーカー／製品名: ChIPAb+; Upstate^®
• テクニック: ChIP: suitable (ChIP-seq); immunoprecipitation (IP): suitable
• アイソタイプ: IgG
• NCBIアクセッション番号: NP_005216
• UniProtアクセッション番号: Q01094
• 輸送温度: dry ice

詳細

詳細

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.

特異性

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

アプリケーション

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.

品質

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.

ターゲットの説明

~55 kDa

物理的形状

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

アナリシスノート

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

法的情報

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

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

安全性情報

• 保管分類コード: 10 - Combustible liquids