S100 Antibody Staining Protocol for Immunohistochemistry

 

Description: S100 belongs to the family of calcium binding proteins such as calmodulin and troponin C. S100A is composed of an alpha and beta chain whereas S100B is composed of two beta chains. Antibody to S100 stains Schwannomas, ependymomas, astrogliomas, almost all benign and malignant melanomas and their metastases. S100 protein is also expressed in the antigen presenting cells such as the Langerhans cells in skin and interdigitating reticulum cells in the paracortex of lymph nodes.

 

Primary Antibody

Name: S100 (Ab-2) Antibody

Clone: Rabbit polyclonal

Supplier: Lab Vision

Catalog Number: RB-044-A

Dilution: 1:800 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

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
Staining Pattern: Cytoplasmic
Images: Search image

Additional Information:
Species Reactivity: Human, mouse, rat, chicken, cow, kangaroo
Fixation: Formalin fixed paraffin sections. Not suitable for frozen 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

 

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