Ordering Information
- : FCCS100182
- : 100 Tests / Kit
Product Images
HeLa cells were treated with 100 µM of etoposide and then stained with Anti-phospho-Histone H2A.X, Alexa Fluor® 488 antibody. As seen in (A), a notic...
HeLa cells were pre-treated with various small molecule inhibitors for 24 hours prior to staining and acquisition. EC50 curves are derived from half ...
FlowCellect™ Histone H2A.X Phosphorylation Assay Kit
Description:
FlowCellect™ Histone H2A.X Phosphorylation Assay Kit
Trade Name:
FlowCellect
Qty/Pk:
100 Tests / Kit
Product Overview:
Millipore’s FlowCellect™ Histone H2A.X Phosphorylation Assay kit includes one directly conjugated antibody, a phospho-specific Anti-phospho-Histone H2A.X (Ser139)-Alexa Fluor®488 conjugated antibody to measure levels of Histone H2A.X. This kit is designed to detect the extent of Histone H2A.X pathway activation by measuring H2A.X phosphorylation in any given cell population. The kit provides a sensitive and valuable tool to study the factors that induce DNA damage and/or affect DNA repair, and allow one to explore the linkage between DNA damage, cell cycle checkpoints, and initiation of apoptosis.
Ionizing radiation (IR) and many chemotherapeutic agents like etoposide kill cancer cells by induction of DNA DSBs. Several reports show that the level of gamma-H2A.X as detected by flow cytometry correlates with the number of DNA strand breaks, to the level of cell death and radiosensitivity. H2A.X phosphorylates in response to a DNA damaging reagent (e.g. Etoposide) or UV light, and its activation clearly indicates that DSBs have occurred. Understanding when DSBs take place can help researchers understand the mechanisms involved in DNA repair and the DNA damage response.
The antibody provided in the kit has been carefully titrated to ensure the ability to measure phospho-Histone H2A.X for accurate determination of protein activation. Sufficient reagents are provided to perform 100 tests. Detailed assay instructions are included to assist in analysis and to ensure the correct cell concentration is obtained during acquisition of sample data.
Ionizing radiation (IR) and many chemotherapeutic agents like etoposide kill cancer cells by induction of DNA DSBs. Several reports show that the level of gamma-H2A.X as detected by flow cytometry correlates with the number of DNA strand breaks, to the level of cell death and radiosensitivity. H2A.X phosphorylates in response to a DNA damaging reagent (e.g. Etoposide) or UV light, and its activation clearly indicates that DSBs have occurred. Understanding when DSBs take place can help researchers understand the mechanisms involved in DNA repair and the DNA damage response.
The antibody provided in the kit has been carefully titrated to ensure the ability to measure phospho-Histone H2A.X for accurate determination of protein activation. Sufficient reagents are provided to perform 100 tests. Detailed assay instructions are included to assist in analysis and to ensure the correct cell concentration is obtained during acquisition of sample data.
Background Information:
H2A.X is a member of the histone H2A family. Histone H2A.X resides downstream of the DNA damage kinase signaling cascade. Phosphorylation of Histone H2A.X at serine 139 is an important indicator of DNA damage. As the level of DNA damage increases, the level of phospho Histone H2A.X (also known as γH2AX) increases, accumulating at the sites of DNA damage. This accumulation of phospho Histone H2A.X is often used to indicate the level of DNA damage present within the cell. H2AX is also responsible for recruiting response proteins to the site of DNA damage and may play a role in DNA repair.
In all, a comprehensive understanding of Histone H2A.X activity and understanding the consequence of its activation can provide the researcher with useful information which will be important in understanding the intrinsic nature of the DNA damage response. Millipore’s FlowCellect Histone H2A.X Phosphorylation Assay Kit is designed to allow the researcher to monitor and accurately measure phospho-specific Histone H2A.X activation in a population of cells.
All FlowCellect kits are optimized on guava® bench top flow cytometers. FlowCellect kits can be used on any flow cytometer following the same protocol providing researchers a reliable and fully validated solution to study the Histone H2A.X signaling pathway right in the comfort of their own lab. The directly conjugated antibody provided in the kit has been carefully titrated and optimized to ensure maximal performance alleviating the need for any additional optimization. This kit contains optimized fixation, permeabilization, wash, and assay buffers to provide researchers with a complete solution for Histone H2A.X signaling analysis.
In all, a comprehensive understanding of Histone H2A.X activity and understanding the consequence of its activation can provide the researcher with useful information which will be important in understanding the intrinsic nature of the DNA damage response. Millipore’s FlowCellect Histone H2A.X Phosphorylation Assay Kit is designed to allow the researcher to monitor and accurately measure phospho-specific Histone H2A.X activation in a population of cells.
All FlowCellect kits are optimized on guava® bench top flow cytometers. FlowCellect kits can be used on any flow cytometer following the same protocol providing researchers a reliable and fully validated solution to study the Histone H2A.X signaling pathway right in the comfort of their own lab. The directly conjugated antibody provided in the kit has been carefully titrated and optimized to ensure maximal performance alleviating the need for any additional optimization. This kit contains optimized fixation, permeabilization, wash, and assay buffers to provide researchers with a complete solution for Histone H2A.X signaling analysis.
Key Applications:
Flow Cytometry
Species Reactivity:
Human
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.
Quality Assurance:
Lot to lot in-process testing on all kit components
Components:
- 1. 20X Anti-Histone H2A.X-Alexa Fluor® 488: (Part No. CS208216) One vial containing 600 µL.
- 2. Fixation Buffer: (Part No. CS202122) One bottle containing 13 mL.
- 3. 10X Wash Buffer: (Part No. CS202123) One bottle containing 13 mL.
- 4. 5X Assay Buffer: (Part No. CS202124) One 55 mL bottle
- 5. 1X Permeabilization Buffer: (Part No. CS203284) One bottle containing 14 mL.
Host:
Mouse
Storage Conditions:
4-8°C for antibodies and buffers
Detection Methods:
Fluorescent
Product Name:
FlowCellect™ Histone H2A.X Phosphorylation Assay Kit
Alternate Names:
Flow Cellect
Flow Cytometry Software Type:
ExpressPro & inCyte
Antibody Type:
Monoclonal Antibody
Flow Cytometry Market:
Research