Bcl-3 Oncoprotein Antibody Staining Protocol for Immunohistochemistry


Description: Bcl-3 was first identified as a putative proto-oncogene and was originally isolated though its involvement in the translocation t(14;19). The Bcl-3 gene product is also thought to play a role in the immune system through its interactions with the NF-kappaB family of transcription factors, to enhance proliferation and to act as a transcription cofactor. More specifically, Bcl-3 appears to regulate the activity of homodimeric NF-kappaB p50 subunit and a closely-related homolog, p52, in a phosphorylation-dependent manner. Bcl-3 mRNA is found in a number of tissues, including spleen and other lymphoid tissues.


Primary Antibody

Name: Bcl-3 Oncoprotein Antibody

Clone: 1E8, Mouse Anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-Bcl-3

Dilution: 1:100 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: Nulcear
Images: Search image

Additional Information:
Tissue Type: Tonsil
Fixation: Formalin-fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Tonsil
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


There is no need to perform antigen retrieval on acetone fixed frozen sections