Bcl-rambo Antibody Staining Protocol for Immunohistochemistry


Description: Bcl-rambo is a member of the Bcl-2 family of proteins which plays a central regulatory role in apoptosis. In mammalian cells, Bcl-rambo is localised to mitochondria where its overexpression induces apoptosis. Bcl-rambo cell death activity is induced through its membrane-anchored C-terminal domain and not by the Bcl-2 homologous region. This C-terminal membrane anchor and its unique extension region is important for pro-apoptotic activity inducing caspase-3 activation. Bcl-rambo mRNA is reported to be found in a diverse range of tissues, with the highest levels detected in heart, pancreas, placenta and skeletal muscle. Bcl-rambo mRNA has also been found in brain, liver and kidney.


Primary Antibody

Name: Bcl-rambo Antibody

Clone: 13E6, Mouse Anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-Bcl-rambo

Dilution: 1:25 - 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: Granular Cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Testis or myocardium
Fixation: Formalin-fixed paraffin sections
Positive Control: Testis or myocardium
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