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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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