 |
|
Bcl-w Antibody Staining Protocol for Immunohistochemistry
Description: Bcl-w belongs to the bcl-2 family of proteins and promotes cell survival, whereas other members such as bak and bax are antagonists and promote apoptosis. The Bcl-w gene is highly conserved between mice and man. Bcl-w protein is found in a diverse range of tissues
including cerebellum, hippocampus, colon, liver, heart, stomach, skeletal muscle, testis and
placenta. It is also expressed in B and T cell lines. Bcl-w is apparently dispensable in normal
development and function of most organs but is essential for spermatogenesis.
Primary Antibody
|
Name: Bcl-w Antibody |
|
Clone: 6C1, Mouse Anti-Human |
|
Supplier: Novocastra Labs |
|
Catalog Number: NCL-Bcl-W |
|
Dilution: 1:20 - 1:40 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: 60min/room temperature |
Antigen Retrieval
| 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 |
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: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
| Tissue Type: Cerebellum |
| Fixation: Formalin-fixed paraffin sections |
| Positive Control: Cerebellum |
| 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: