S100 Antibody Staining Protocol for Immunohistochemistry


Description: S100s are a unique group of EF-hand proteins characterized by cell-type and cell cycle- specific expression, as well as deregulated expression in neurological disorders (S100B- Alzheimer disease, Down syndrome, and epilepsy), inflammatory disorders (S100A8/A9- cystic fibrosis, arthritis and chronic bronchitis), and certain cancers (S100A2/A4/A6). At the cellular level, S100 proteins have been implicated in the control of cell growth and proliferation, cell cycle progression, and modulation of specific signal transduction pathways, but also have extracellular functions, including neurotrophic and antimicrobial activity. S-100 protein derived from brain tissue is an acidic calcium-binding protein with molecular weight of about 21kda. In human brain tissue, S-100 protein is mainly presented by two isoforms, a Beta-Beta(BB) homodimer or S-100b and an Alpha-Beta (AB) heterodimer or S-100a. Becasue of its predomiant location in astrological cells, S-100 protein can be used as a sensitive and reliable marker for central nervous system injury. Structural damage of glial cells causes leakage of S-100 protein into extracellular matrix and into cerebrospinal fluid, further releasing into the bloodstream. Measurements of S-100 protein in patient serum samples maybe useful in monitoring of traumatic brain injury, ischemic brain damage after circulatory arrests, in diagnosis and prognosis of clinical outcome in acute stroke.


Primary Antibody

Name: S100 Antibody

Clone: Rabbit anti-Human

Supplier: Dako

Catalog Number: Z0311

Dilution: 1:1000 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: No antigen retrieval needed
Buffer/pH value: N/A
Heat/Cool Temperature: N/A
Heat/Cool Time: N/A

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 Reactivity: Human
Fixation: Formalin fixed paraffin sections
Positive Control: Melanoma, Schwannomas
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