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  • 17-10254 - ChIPAb+ Histone H3 (CT) - ChIP Validated Antibody and Primer Set

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17-10254 Sigma-Aldrich

ChIPAb+ Histone H3 (CT) - ChIP Validated Antibody and Primer Set

Synonym: Histone H3.1t, H3t, histone 3, H3/g

  •  eCl@ss 32160702

  •  NACRES NA.32

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Properties

Related Categories Antibodies, Antibodies for Epigenetics and Nuclear Signaling, ChIP Validated Antibodies, Primary Antibodies
biological source   rabbit
antibody form   serum
clone   polyclonal
species reactivity   yeast, mouse, chicken, chicken, mouse, human, mouse, human, yeast, rat, rat, Saccharomyces cerevisiae, chicken, human, rat
mfr. no.   ChIPAb+
  Upstate®
application(s)   ChIP: suitable
  dot blot: suitable
  immunocytochemistry: suitable
  inhibition assay: suitable (peptide)
  western blot: suitable
NCBI accession no.   NM_002107.3
UniProt accession no.   Q16695
shipped in   dry ice

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 H3 (C-Term) set includes the Histone H3 (C-Term) antibody, a Normal Rabbit Serum, and control primers which amplify a 110 bp region of ChIP Primers, human β-globin. The Histone H3 (C-Term) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Histone H3 (C-Term)-associated chromatin.

Specificity

Broad species cross-reactivity expected due to sequence homology.

Recognizes C-Terminal region of Histone H3, Mr 17 kDa

Immunogen

KLH-conjugated, synthetic peptide corresponding to the C-terminus of human Histone H3.

Application

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 Rabbit Serum,or 1 µL of Anti-Histone H3 (C-Term) and the Magna ChIP A Kit (Cat. # 17-610). Successful immunoprecipitation of histone H3 associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin as a positive locus, and GAPDH promoter primers (22-004) as a negative locus. (Figure 2). 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.
Acid extracted proteins from HeLa cells untreated (lane 1) or treated with sodium butyrate (lane 2) or colcemid (lane 3) and recombinant Histone H3 (lane 4) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Histone H3, CT, pan (1:50,000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Histone H3 (~17 kDa). (Figure 3).

Packaging

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

Quality

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 Rabbit Serum, or 1 µL of Anti-Histone H3 (C-Term) and the Magna ChIP® A Kit (Cat. # 17-610). Successful immunoprecipitation of histone H3 associated DNA fragments was verified by qPCR using ChIP Primers, human β-globin (Figure 1).
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Target description

Approx. 17 kDa

Physical form

Anti-Histone H3 (C-Term) (rabbit polyclonal). One vial containing 25 µL of rabbit antiserum diluted 1:2 in storage buffer (0.02 M Phosphate, 0.25 M NaCl, 0.1% sodium azide) before the addition of glycerol to 30%. Store at -20°C.
Normal Rabbit Serum. One vial containing 25 μL of antiserum containing 0.05% sodium azide. Store at -20°C.
ChIP Primers, human 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

Analysis Note

Control
Includes normal rabbit serum and primers specific for human β-globin.

Legal Information

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

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.

Documents

Certificate of Analysis (COA)

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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 ...
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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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