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

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: 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

 

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