Pax-5 Antibody Staining Protocol for Immunohistochemistry


Description: The Pax gene family of nuclear transcription factors is comprised of nine family members that function during embryogenesis to regulate the temporal and position-dependent differentiation of cells. In addition, the family is involved in a variety of signal transduction pathways in the adult organism. Mutations in the Pax family of proteins have been linked to disease and cancer in humans. For example, the human Pax-5 gene encodes a B cell lineage specific protein called B cell specific activator protein or BSAP, which is expressed in pro-B, pre-B and mature B lymphocytes but not in plasma cells. BSAP functions to regulate not only B cell development, but also influences the balance between immunoglobulin secretion and B cell proliferation. Overexpression of BSAP has been implicated in cellular transformation, and in the case of small lyphocytic lymphomas with plasmacytoid differentiation, a t(9;14)(p13;q32) translocation resulting in the deregulation of Pax-5 gene expression has been detected. The gene which encodes Pax-5 maps to human chromosome 9p13.


Primary Antibody

Name: Pax-5 (C-20) Antibody

Clone: Goat Polyclonal

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-1974

Dilution: 1:2000 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: 60 min/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: Nuclear
Images: Search image

Additional Information:
Species Reactivity: Human, mouse, rat
Fixation: Formalin fixed paraffin sections
Positive Control: Tonsil, spleen
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