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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
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Name: Pax-5 (C-20) Antibody |
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Clone: Goat Polyclonal |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-1974 |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
References: