Description: Histone H3 (Mitosis Marker) is specific for phosphorylated histone H3, Mr 17kDa and has been reported to detect mitotic cells in C. elegans and Drosophila embryos by immunohistochemistry.
Primary Antibody
Name: phospho-Histone H3 Antibody |
Clone: Rabbit anti-Human |
Supplier: Upstate |
Catalog Number: 06-570 |
Dilution: 1:200 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, cervix, skin, Alzheimer's brain, colonic adenocarcinoma, carcinoma. |
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:
1. Ogawa O, et al (2003) Ectopic localization of phosphorylated histone H3 in Alzheimer's diseases: a mitotic catastrophe? Acta Neuropathol 105:524-528.
2. Scott IS, et al (2003) A novel immunohistochemical method to estimate cell-cycle phase distribution in archival tissue: implications for the prediction of outcome in colorectal cancer. J Pathology 201:187-197.