Bax Antibody Staining Protocol for Immunohistochemistry


Description: The Bcl-2 gene was isolated at the chromosomal breakpoint of t (14;18)-bearing follicular B cell lymphomas. Bcl-2 blocks cell death following a variety of stimuli and confers a death-sparing effect to certain hematopoietic cell lines following growth factor withdrawal. Bcl-2 is localized to outer mitochondrial membranes and endoplasmic reticulum as well as nuclear membranes. A related protein, designated Bax p21 (for Bcl-associated X protein), has extensive amino acid homology with Bcl-2 and both homodimerizes and forms heterodimers with Bcl-2. Overexpression of Bax accelerates apoptotic death induced by cytokine deprivation in an IL-3 dependent cell line and Bax also counters the death repressor activity of Bcl-2.


Primary Antibody

Name: BAX Antibody

Clone: Rabbit polyclonal

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-526

Dilution: 1:200 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: Breast carcinoma
Fixation: Formalin-fixed paraffin sections
Positive Control: Breast carcinoma
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