Description: The assessment of cell proliferation by the detection of Ki67 antigen in neoplastic cell populations has been shown to be of prognostic value.
Primary Antibody
Name: Ki67 Antibody |
Clone: MIB-1, Mouse anti-Human |
Supplier: Dako |
Catalog Number: M7240 |
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 min/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: 95-100 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 |
Species Reactivity: Human |
Fixation: Formalin fixed paraffin sections or acetone-fixed, frozen sections as well as cell smears. |
Positive Control: Tonsil, thymus, stomach, small intestine, colon, skin, oral mucosa and breast carcinoma, bone marrow, testis, and uterus. |
Negative Control: Omit primary antibody, isotype control, 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 |
References:
1. Charafe-Jauffret E, et al (2004) Immunophenotypic analysis of inflammatory breast cancers: identification of an inflammatory signature. J Pathology. 202:265-273.
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.
3. Heuschmid M, Hofmockel G, Dexler B, Dammrich J, Bassukas ID. (2002) Different antigen unmasking techniques lead to significant differences in immunohistochemical staining of Ki-67 (Mib-1) in renal cell carcinomas. Oncol Rep. Jan-Feb;9(1):19-22. PubMed Abstract
4. Boon ME (1996) Microwave-antigen retrieval: the importance of pH of the retrieval solution for MIB-1 staining Eur J Morphol. 34(5):375-9. PubMed Abstract