17-10141
ChIPAb+ Phospho-Histone H3 (Thr3) - ChIP Validated Antibody and Primer Set, rabbit monoclonal
culture supernatant, clone JY325, from rabbit
Synonym(s):
H3T3P, Histone H3 (phospho T3)
About This Item
Related Categories
General description
The ChIPAb+ Phospho-Histone H3 (Thr3) set includes the Phospho-Histone H3 (Thr3) antibody, a negative control supernatant, and control primers which amplify a 166 bp region of human GAPDH promoter. The Phospho-Histone H3 (Thr3) and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Phospho-Histone H3 (Thr3)-associated chromatin.
The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.
Specificity
Immunogen
Application
Representative lot data.
Sonicated chromatin prepared from untreated or colcemid-treated HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Negative Control Supernatant, or 2 µL ofAnti-Phospho-Histone H3 (Thr3) and the Magna ChIP® A Kit (Cat. # 17-610). Successful immunoprecipitation of Phospho-Histone H3 (Thr3) associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter Region for untreated and treated chromatin samples. 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 histone H3 (Lane 1) and acid extracts from untreated (Lane 2) or colcemid treated (Lane 3) HeLa cells were resolved by
electrophoresis, transferred to nitrocellulose and probed with anti-Phospho-Histone H3 (Thr3), clone JY325 (10 ng/mL). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates Phospho- Histone H3 (~17 kDa).
Dot Blot:
Representative lot data.
A dilution series of Histone H3 peptides containing the following modifications was made:
Column 1: Phospho-threonine 3
Column 2: unmodified, containing Thr3
Column 3: Phospho-threonine 6
Column 4: Phospho-threonine 11
Column 5: Phospho-threonine 22
Column 6: Phospho-serine 10
Column 7: Phospho-threonine 80
Column 8: Phospho-serine 28
2 μL of each dilution was spotted onto PVDF membranes and probed with Anti-Phospho- Histone H3 (Thr3), clone JY325 (20 ng/mL)
Peptides were visualized using a Goat Anti-Rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system..
Epigenetics & Nuclear Function
Histones
Packaging
Quality
Representative lot data.
Sonicated chromatin prepared from colcemid-treated HeLa cells (1 X 10E6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µL of either Negative Control Supernatant, or 2 µL Anti-Phospho-Histone H3 (Thr3) and the Magna ChIP® A Kit (Cat. # 17-610). Successful immunoprecipitation of Phospho-Histone H3 (Thr3) associated DNA fragments was verified by qPCR using ChIP Primers, human GAPDH promoter Region.
Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.
Target description
Physical form
Negative Control Supernatant. One vial containing 100 µL of cultured supernatant in 0.05% sodium azide. Store at -20°C.
Control Primers, human GAPDH promoter. 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
Analysis Note
Includes negative control supernatant and primers specific for human GAPDH promoter.
Legal Information
Disclaimer
Storage Class Code
10 - Combustible liquids
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PDSCL
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FSL
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ISHL Indicated Name
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ISHL Notified Names
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Cartagena Act
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JAN Code
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