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  • 17-10048 - ChIPAb+ Histone H2A.Z - ChIP Validated Antibody and Primer Set

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17-10048 Sigma-Aldrich

ChIPAb+ Histone H2A.Z - ChIP Validated Antibody and Primer Set

Synonym: H2A histone family, member Z, H2AZ histone

  •  eCl@ss 32160702

  •  NACRES NA.75

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Properties

Related Categories Antibodies, Antibodies for Epigenetics and Nuclear Signaling, ChIP Validated Antibodies, Primary Antibodies
clone   polyclonal
biological source   sheep
application(s)   ChIP: suitable
  immunoprecipitation (IP): suitable
  western blot: suitable
species reactivity   human, human
shipped in   dry ice
purified by   affinity chromatography
mfr. no.   ChIPAb+
  Upstate®
NCBI accession no.   NP_036544
UniProt accession no.   P0C0S5
Gene Information   human ... H2AFZ(3015)

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 H2A.Z set includes the Histone H2A.Z antibody, a negative control antibody (normal sheep IgG), and qPCR primers which amplify a 166 bp region of human GAPDH. The Histone H2A.Z and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H2A.Z -associated chromatin.

H2A.Z, also known as H2AFZ, is a variant of histone H2A variant that is highly conserved from yeast to humans. Like other histone variants, H2A.Z is encoded by a unique gene, resulting in a slightly divergent type of H2A with specialized functions. H2A.Z is involved in chromatin compaction by stabilizing the histone octamer within the nucleosome. It promotes gene expression through the creation of an open domain of chromatin that is more easily transcribed. H2A.Z is nonrandomly distributed throughout the genome, targeted primarily to pericentric heterochromatin, but excluded from the inactive X chromosome and the nucleolus. H2A.Z is associated with both euchromatin and facultative heterochromatin. DNA methylation can influence chromatin structure and effect gene silencing by excluding H2A.Z, and likewise, H2A.Z protects genes from DNA methylation. Thus, H2A.Z plays a role in regulating heterochromatin silencing and is involved in transcriptional control.

Specificity

Predicted to cross-react with zebrafish, mouse, chicken, rat, rabbit, and Xenopus based on sequence homology.

Recognizes Histone H2A.Z, Mr ~15 kDa.

Wide species reactivity is expected.

Immunogen

Epitope: C-terminus

Linear peptide corresponding the C-terminal amino acids 113-127 of human Histone H2A.Z.

Application

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Sheep IgG or 1 µg of Anti-Histone H2A.Z and the Magna ChIP G Kit (Cat. # 17-611).
Successful immunoprecipitation of Histone H2A.Z associated DNA fragments was verified by qPCR using Control Primers (Part No. 22-004) for a positive, and GAPDH Coding 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
HeLa lysate was resolved by electrophoresis, transferred to PVDF and probed with anti-Histone H2A.Z (1:1000 dilution).
Proteins were visualized using a rabbit anti-sheep secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Histones

Target description

~15 kDa

Physical form

Affinity Purfied

Anti-Histone H2A.Z (Sheep polyclonal IgG). One vial containing 25 μg of affinity purified sheep polyclonal IgG in PBS, pH 7.4 containing 1% BSA with 0.02% sodium azide, before the addition of 30% glycerol. Store at -20°C.

Normal Sheep IgG. One vial containing 25 µg of protein G purified sheep IgG in 0.02 M potassium phosphate, 0.15 M NaCl, pH 7.2, with 0.01% sodium azide, before the addition of 30% glycerol. Store at -20°C.

Control Primers. One vial containing 75 μL of 5 μM of each primer specific for human GAPDH. Store at -20°C.
FOR: TAC TAG CGG TTT TAC GGG CG
REV: TCG AAC AGG AGG AGC AGA GAG CGA

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: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

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.

Legal Information

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

Packaging

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

Quality

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Sheep IgG or 1 µg of Anti-Histone H2A.Z and the Magna ChIP® G Kit (Cat. # 17-611). Successful immunoprecipitation of H2A.Z associated DNA fragments was verified by qPCR using Control Primers (Please see figures).

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

Analysis Note

Control
Includes negative control sheep IgG antibody and primers specific for human GAPDH.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.

Documents

Certificate of Analysis (COA)

Please Enter a Lot Number
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detection methods, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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