 Methods & Techniques For Histologists and Immunohistochemists |
BAG-1 (RAP46 Protein) Antibody Staining Protocol for Immunohistochemistry
Description: BAG-1, also called RAP46, is a multifunctional protein that regulates several cellular processes including cell survival, signal transduction and protein folding. BAG-1 is an anti-apoptotic protein capable of interacting with several types of proteins including those of the bcl-2 family, Raf-1, certain tyrosine kinase growth factor receptors and steroid hormone receptors. Alternative translation initiation points in the BAG-1 transcript results in the expression of four protein isoforms known as p50, p46, p33 and p29. The protein is expressed in a number of cell lines and has been demonstrated immunohistochemically to stain both the cytoplasm and nuclei of normal breast tissue.
Primary Antibody
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Name: BAG-1 (RAP46 Protein) Antibody |
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Clone: 5C5, Mouse Anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-BAG-1 |
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Dilution: 1:25 - 1:50 |
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Incubation Time/Temp: 60min/room temperature |
Antigen Retrieval
| Device: Steamer |
| Buffer/pH value: Citrate buffer/pH 6.0 |
| 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: Nuclear and cytoplasmic |
| Images: Search image |
Additional Information:
| Tissue Type: Tonsil |
| Fixation: Formalin-fixed paraffin sections |
| Positive Control: Tonsil |
| 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: