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S-100 Protein Antibody Staining Protocol for
Immunohistochemistry
Description: S-100 protein is a member of the EF-hand calcium-binding protein superfamily that is widely distributed and conserved in the central nervous system of vertebrates. The protein exists in both heterodimeric and homodimeric forms. The two subunits of S-100 protein are the products of separate genes that are differentially expressed by various cells. The beta subunit is present in all S-100 positive cells and tumors. In contrast, the alpha subunit is detectable only in heart, skeletal muscle and brain. In addition to its calcium binding properties, S-100 protein has a high-affinity binding site for zinc and is involved in the regulation of protein phosphorylation in the brain. The protein apparently plays a role in cell differentiation and growth, cytoskeletal structure and function, and has been implicated in neuropathological diseases.
Primary Antibody
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Name: S-100 Protein Antibody |
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Clone: Mouse Anti-S-100 Protein |
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Supplier: Millipore |
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Catalog Number: MAB079-1 |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
| Species Activity: Bovine, canine, human, mouse, rat, rabbit, feline. |
| Tissue Type: Skin |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Melanoma |
| Negative Control: Omit primary antibody, isotype control or 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: