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Synthetic phospho-peptide, which represents a portion of the C-terminus of human histone H2AX surrounding phosphorylated serine 139 (LocusLink ID 3014).
Our Abpromise guarantee covers the use of ab11174 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent dilution.|
|WB||1/5000 - 1/15000. Detects a band of approximately 15 kDa.|
|Flow Cyt||Use 0.5µg for 106 cells.|
|ICC||1/400 - 1/800.|
Samples: Nuclear extract (50
UT = untreated
Ub = ubiquitylated
NCS = neocarzinostatin - 200 ng/ml, 30 min
ab11174 at 1/1000 staining human HeLa cells by ICC/IF. These cells express a gene that causes a DNA damage response, leading to H2AX phosphorylation. The cells were paraformaldehyde fixed and blocked with BSA prior to incubation with the antibody for 45 minutes. An Alexa-Fluor ® 488 conjugated goat anti-rabbit was used as the secondary.
ab11174 at 0.5ug staining gamma H2A.X in human Jurkat cell line by flow cytometery. Cells were treated with 5ug/ml etopside for 3 hours, fixed in 1.5% paraformaldehyde, and permeabilized in 90% methanol. A FITC-conjugated rabbit polyclonal was used as secondary (in 150ul reaction). The black line indicates cells treated with etopside and anti KLH antibody, the red line repersents untreated cells and anti gamma H2A.X and blue line indicates etopside treated anti gamma H2A.X respectively.
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