ChIPAb + Histone H2B

Non disponible en dehors du Royaume-Uni et de l'Irlande

Analysis Note

ControlIncludes negative control normal rabbit IgG antibody and primers specific for human β-globin.

Application

Research Sub CategoryHistones

The ChIPAb+ Histone H2B set includes the Histone H2B antibody, a negative control rabbit IgG & qPCR primers which amplify a 110 bp region of human β-globin promoter.

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10⁶ cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µL of Normal Rabbit IgG or 1 µL of Anti-Histone H2B and the Magna ChIP™ A Kit (Cat. # 17-610).
Successful immunoprecipitation of Histone H2B associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin for a positive, and human GAPDH primers as a negative assay (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.
Recombinant Xenopus Histone H2B, chicken core histones and acid extract from either sodium butyrate or colcemid treated HeLa cells were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Histone H2B (1:5,000 dilution).
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).

Research CategoryEpigenetics & Nuclear Function

Disclaimer

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.

General description

Histone H2B is one of the 5 main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N terminal tail H2A is involved with the structure of the nucleosomes of the ′beads on a string′ structure.

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 H2B set includes the Histone H2B antibody, a negative control rabbit IgG, and qPCR primers which amplify a 110 bp region of human β-globin promoter. The Histone H2B and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H2B-associated chromatin.

Immunogen

KLH-conjugated, synthetic peptide corresponding to amino acids 118-126 (CG-AVTKYTSSK) of human Histone H2B, with an N-terminal CG added for conjugation purposes.

Legal Information

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

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

Packaging

25 assays per set. Recommended use: ~1 µL of antibody per chromatin immunoprecipitation (dependent upon biological context).

Physical form

Anti-Histone H2B (rabbit polyclonal). One vial containing 25 µL of protein A purified rabbit polyclonal in buffer containing 0.02 M phosphate buffer, pH 7.6, 0.25 M NaCl, and 0.1% sodium azide, and 30% glycerol. Store at -20°C.

Normal Rabbit IgG. One vial containing 125 µg of purified rabbit IgG in 125 µL storage buffer. Store at -20°C.

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

Protein A purified

Format: Purified

Quality

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 10⁶ cell equivalents per IP) were subjected to chromatin immunoprecipitation using either 1 µL of Normal Rabbit IgG or 1 µL of Anti-Histone H2B and the Magna ChIP^® A Kit (Cat. # 17-610).
Successful immunoprecipitation of Histone H2B associated DNA fragments was verified by qPCR using ChIP Primers, human 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.

Specificity

Recognizes and is specific for Histone H2B, MW ~15 kDa. An additional band at ~23 kDa is detected in some samples, which likely corresponds to ubiquityl-Histone H2B.

Others not tested.

Broad species cross-reactivity expected based on sequence homology.

Storage and Stability

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variabillity in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Target description

~15 kDa