Phospho-Histone H3(Ser 10) Assay
(Mouse primary antibody)
Description:
Phospho-Histone H3(Ser 10) Assay
(Mouse primary antibody)
(Mouse primary antibody)
Product Overview:
Millipore's HCS203 assay provides a complete solution for identifying and quantifying phospho-histone H3(Ser10) expression in cellular imaging applications. The reagents in the kit have been specifically optimized for HCS applications.
Background Information:
Histone H3 has been identified as a crucial target for cancer chemotherapy due to its important regulatory role in cell proliferation and neoplastic cell transformation. Accordingly, HCS of histone H3 phosphorylation is increasingly being used as a tool in drug discovery. HCS assays quantifying phospho-histone H3 levels have recently been utilized to characterize Aurora kinase inhibitors. Additionally, histone H3 phosphorylation has been used as a readout in cell cycle inhibitor profiling studies, illustrating the value of this approach in drug discovery applications.
Key Applications:
in vitro Toxicology Assays
Application Notes:
Cell Cycle
Usage Statement:
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.
Therapeutic Areas:
Oncology
Antigen:
Phospho-histone H3
Host Cells:
HeLa cell line
Materials Required but Not Delivered:
1. Sterile, tissue culture-treated black/clear bottom microplates suitable for High-Content Imaging.
2. Cell-type for assay, e.g., HeLa (human cervical adenocarcinoma, ATCC #CCL-2) or A549 (human lung carcinoma, ATCC #CCL-185).
3. Tissue culture instruments/supplies (including 37ºC incubator, growth media, flasks/plates, detachment buffer, etc.) for cell type of interest.
4. HCS imaging/analysis system, e.g., GE Healthcare IN Cell Analyzer 1000 with Investigator software. Imaging system must be equipped with beam-splitters and filters capable of reading emission spectra in the blue and red ranges.
2. Cell-type for assay, e.g., HeLa (human cervical adenocarcinoma, ATCC #CCL-2) or A549 (human lung carcinoma, ATCC #CCL-185).
3. Tissue culture instruments/supplies (including 37ºC incubator, growth media, flasks/plates, detachment buffer, etc.) for cell type of interest.
4. HCS imaging/analysis system, e.g., GE Healthcare IN Cell Analyzer 1000 with Investigator software. Imaging system must be equipped with beam-splitters and filters capable of reading emission spectra in the blue and red ranges.
Components:
- 1. Mouse Anti-Phospho-Histone H3(Ser10) HCS Primary Antibody, 100X
- 2. HCS Secondary Antibody (donkey anti-mouse IgG, Cy3 conjugate), 200X
- 3. Hoechst HCS Nuclear Stain, 200X
- 4. HCS Fixation Solution with Phenol Red, 2X
- 5. HCS Immunofluorescence Buffer, 1X
- 6. HCS Wash Buffer, 1X
- 7. Etoposide (S/G2 Arrest Control Compound), 250X
- 8. Vinblastine Sulfate (G2/M Arrest Control Compound), 250X
- 9. DMSO for Compound Serial Dilution
- 10. Compound Dilution Buffer
- 11. Plate Sealers
Kit Type:
High-Content Screening
Configuration:
Sufficient reagents for 5 X 96 well plates
Detection Methods:
Fluorescent
Storage Conditions:
When stored under the conditions indicated on the labels, the kit components are stable up to the expiration date. HCS Fixation Solution, HCS Immunofluorescence Buffer, HCS Wash Buffer, DMSO, Compound Dilution Buffer, and Plate Sealers should be stored at 2-8ºC. HCS Primary Antibody, HCS Secondary Antibody, Hoechst HCS Nuclear Stain, Etoposide and Vinblastine Sulfate should be stored at -20ºC, avoiding repeated freeze/thaw cycles. (Note: If kit is expected to be used for multiple experiments, rather than a single use, thaw antibodies, nuclear stain and control compounds and dispense into appropriately sized aliquots. Store aliquots at -20ºC.)
Discard any remaining reagents after the expiration date.
Discard any remaining reagents after the expiration date.
Host:
Mouse