 Methods & Techniques For Histologists and Immunohistochemists |
Bcl-6 Antibody Staining Protocol for Immunohistochemistry
Description: Bcl6 is a proto-oncogene that encodes Kruppel type Zinc finger protein of 95 kDa and shares homology with transcription factor. Bcl6 is mainly expressed in normal germinal center B cells and related lymphomas. Bcl6 plays an important role in the regulation of the Th2 inflammatory response. Mice deficient in Bcl6 can develop severe Th2-type inflammation in multiple organs. This antibody reacts with Bcl-6 gene product in follicular lymphomas, diffuse large B cell lymphomas, Burkitt's lymphomas and in nodular, lymphocyte predominant Hodgkin's disease.
Primary Antibody
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Name: Bcl-6 Antibody |
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Clone: P1F6, Mouse Anti-Human |
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Supplier: Abcam |
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Catalog Number: ab9479 |
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Dilution: 1:20 |
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Incubation Time/Temp: 60min/room temperature |
Antigen Retrieval
| Device: Steamer |
| Buffer/pH value: EDTA/pH 8.0 |
| Heat/Cool Temperature: 95-100 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods |
Chromogen Substrate
| Reagent: DAB |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Nulcear |
| Images: Search image |
Additional Information:
| Tissue Type: Tonsil, follicular lymphomas |
| Fixation: Formalin-fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Tonsil, follicular lymphomas |
| 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 |
Notes:
There is no need to perform antigen retrieval on acetone fixed frozen sections |
References: