Bcl-2 Antibody Staining Protocol for Immunohistochemistry


Description: Apoptosis defines a set of cascades which, when initiated, programs the cell to undergo lethal changes such as membrane blebbing, mitochondrial break down and DNA fragmentation. Bcl-2 is one among many key regulators of apoptosis which are essential for proper development, tissue homeostasis and protection against foreign pathogens. Human Bcl-2 is a 26 kDa, membraneassociated, anti-apoptotic oncoprotein that can promote cell survival through protein-protein interactions with other Bcl-2 related family members, such as the death suppressors Bcl-xL, Mcl-1, Bcl-w and A1, or the death agonists Bax, Bak, Bik, Bad and BID. The anti-apoptotic function of Bcl-2 can also be regulated through proteolytic processing and phosphorylation. Bcl-2 may promote cell survival by interfering with the activation of the cytochrome c/Apaf-1 pathway through stabilization of the mitochondrial membrane. Mutations in the Bcl-2 gene can contribute to cancers where normal physiological cell death mechanisms are compromised by deregulation of the antiapoptotic influence of Bcl-2.


Primary Antibody

Name: Bcl-2 (C-2) Antibody

Clone: Mouse monoclonal

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-7382

Dilution: 1:50 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: Normal human colon or prostate
Fixation: Formalin-fixed paraffin sections
Positive Control: Normal human colon or prostate
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