S-100 Protein Antibody Staining Protocol for



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

Name: S-100 Protein Antibody

Clone: Mouse Anti-S-100 Protein

Supplier: Millipore

Catalog Number: MAB079-1

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:
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