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-Tek^TM 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

References: