Description: Histone H2AX is a 14 kDa ubiquitous member of the H2A histone family that contains an evolutionary conserved SQ motif at the C-terminus in eukaryotes. Serine 139 within this motif becomes rapidly phosphorylated to yield a form known as γ-H2AX in response to double-strand DNA damage and apoptosis. Phosphorylation reaches half its maximum between 1-3 minutes after DNA damage occurs, and hundreds to several thousand molecules of γ-H2AX are present per double-strand break. This antibody is unique in only detecting double strand DNA breaks.
Primary Antibody
Name: Histone H2AX Antibody |
Clone: Rabbit polyclonal |
Supplier: Trevigen |
Catalog Number: 4411-PC-100 |
Dilution: 1:100 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
Incubation Time/Temp: 60 minutes/room temperature |
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
Heat/Cool Temperature: 95 ºC - 100 ºC/room temperature |
Heat/Cool Time: 20 minutes/20 minutes |
Standard Method: ABC Method or LSAB Method |
Enhanced Method: Polymeric Methods |
Reagent: DAB |
Incubation Time/Temperature: 1-3 minutes/room temperature |
Reagent: Mayer's Hematoxylin |
Staining Time: 30 seconds |
Staining Pattern: Nuclear |
Images: Search image |
Tissue Type: Tonsil, skin |
Fixation: Formalin fixed paraffin sections |
Positive Control: Tonsil, skin |
Negative Control: Omit primary antibody, isotype control or absorption control |
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
Not tested on frozen sections. |
References: