Bcl-x Antibody Staining Protocol for Immunohistochemistry


Description: bcl-x has homology with and is a member of the bcl-2 family of proteins. bcl-x can function as a regulator of cell death independently of bcl-2. Differential splicing of the bcl-x mRNA produces short and long variants known as bcl-xS and bcl-xL. These variants have different functions. bcl-x immunoreactivity has been demonstrated in many cell types and like bcl-2, has been localised to the cytosol associated with mitochondria. bcl-x has been immunohistochemically detected in plasma cells, activated lymphocytes in interfollicular areas and a small number of lymphocytes within germinal centres. In normal tissues, bcl-x has been reported in cortical thymocytes, as well as spermatocytes and spermatids in the testes. It is also found in mammary epithelial cells, secretory and basal epithelial cells and differentiated keratinocytes in the upper layers of the epidermis (but not in basal cells). In many cases, the pattern of bcl-x expression is markedly different from that reported for bcl-2.


Primary Antibody

Name: Bcl-x Antibody

Clone: NC1, Mouse Anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-Bcl-X

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: Antigen retrieval is not needed
Buffer/pH value: N/A
Heat/Cool Temperature: N/A
Heat/Cool Time: N/A

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: Skin, differentiated keratinocytes
Fixation: Formalin-fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Skin, differentiated keratinocytes
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