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ChIPAb+ Histone H4 - ChIP Validated Antibody and Primer Set, rabbit monoclonal，25ASSAYS，17-10047，Sigma

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订货号： 0BW1766

品牌型号： Sigma 17-10047

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最小订货量： 1件

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商品介绍
规格参数
包装参数

产品介绍 Product Description

General description

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+ Histone H4 set includes the Histone H4 antibody, a negative control antibody (normal rabbit IgG), and qPCR primers which amplify a 110 bp region of human β-Globin. The Histone H4 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H4-associated chromatin.
~11 kDa

Immunogen

Epitope: a.a 25-28
KLH-conjugated synthetic peptide corres-ponding to amino acids 17-28 of Histone H4.

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10⁶ cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either normal rabbit IgG or 2 µL of Anti-Histone H4 antibody and the Magna ChIP A Kit (Cat. # 17-610).
Successful immunoprecipitation of total Histone H4 associated DNA fragments was verified by qPCR using ChIP Primers, β-Globin, as well as GAPDH promoter Control Primers, (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 A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Representative Lot Data.
HeLa Acid Extract (Lane 1 and 3), HeLa Acid Extract from sodium butyrate treated cells (Lane 2), HeLa Acid Extract from colcemid treated cells (Lane 4) and recombinant histone H4 (Lane 5) were resolved by electrophoresis, transferred to PVDF membranes and probed with Anti-Histone H4 (1:1,000 dilution).
Proteins were visualized using a Donkey anti-rabbit IgG conjugated to HRP and visualized using a chemiluminescence detection system. (Please see figures).

This ChIPAb+ Histone H4 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Biochem/physiol Actions

Wide range of cross-reactivity expected based on sequence homology.

Physical form

Anti-Histone H4 (rabbit monoclonal IgG). One vial containing 50 µL of protein A purified rabbit monoclonal IgG supernatant in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide before addition of glycerol to 40%. Store at -20°C.

Normal Rabbit IgG. One vial containing 75 µg of normal rabbit IgG in 75 µL storage buffer. Store at -20°C.

ChIP Primers, β-Globin. One vial containing 75 μL of 5 μM of each primer specific for human β-Globin. Store at -20°C.
FOR: AGG ACA GGT ACG GCT GTC ATC
REV: TTT ATG CCC AGC CCT GGC TC

Format: Purified

Analysis Note

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10⁶ cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either normal rabbit IgG or 2 µL of Anti-Histone H4 antibody and the Magna ChIP^® A Kit (Cat. #17-610). Successful immunoprecipitation of total Histone H4 associated DNA fragments was verified by qPCR using ChIP Primers, β-Globin (Please see figures).

Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Control
Includes negative control antibody (normal rabbit IgG) and primers specific for human β-Globin.

Legal Information

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

技术参数 Specifications

biological source	rabbit
Quality Segment	100
clone	monoclonal
species reactivity	human
manufacturer/tradename	ChIPAb+,Upstate^®
technique(s)	ChIP: suitable,immunoprecipitation (IP): suitable,western blot: suitable
NCBI accession no.	NP_001029249
UniProt accession no.	P62805
shipped in	dry ice

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