17-10254
ChIPAb+ Histone H3 (CT) - ChIP Validated Antibody and Primer Set
serum, from rabbit
Synonym(s):
Histone H3.1t, H3t, histone 3, H3/g
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32
biological source
rabbit
Quality Level
antibody form
serum
clone
polyclonal
species reactivity
rat, chicken, mouse, human, Saccharomyces cerevisiae, yeast
manufacturer/tradename
ChIPAb+
Upstate®
technique(s)
ChIP: suitable
dot blot: suitable
immunocytochemistry: suitable
inhibition assay: suitable (peptide)
western blot: suitable
NCBI accession no.
UniProt accession no.
shipped in
dry ice
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.
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).
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).
This ChIPAb+ Histone H3 (C-Term) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
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.
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
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.
Includes normal rabbit serum 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
Storage Class Code
10 - Combustible liquids
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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