ChIPAb+ Phospho-Histone H3 (Thr3)

ChIPAb+ Phospho-Histone H3 (Thr3)

Stock Code: 3586335
Manufacturer Part No: 17-10141
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Analysis Note


ControlIncludes negative control supernatant and primers specific for human GAPDH promoter.


Application


This ChIPAb+ Phospho-Histone H3 (Thr3) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.


Research Sub CategoryHistones


Research CategoryEpigenetics & Nuclear Function


Chromatin Immunoprecipitation:
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..


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 H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the ′beads on a string′ structure.

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

Quality Level100
ManufacturerSIGMA-ALDRICH

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