Fibroblast Surface Protein (FSP) Antibody Staining Protocol for Immunohistochemistry


Description: The antibody recognizes an epitope on primary cultured human fibroblasts, fibroblast cell lines, tissue macrophages and 95% of peripheral blood monocytes. It does not react with vascular smooth muscle cells. The antibody may be used for removal of fibroblasts from human cultured cells by inhibition of fibroblast adherence to the culture vessel or by cytotoxic effect in the presence of rabbit complement.


Primary Antibody

Name: FSP Antibody

Clone: 1B10, Mouse anti-Human

Supplier: Sigma-Aldrich

Catalog Number: F4771

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: 60 minutes/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 ºC - 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: Membrane/cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Human synovial, mammary, foreskin and thymic fibroblasts and in malignant fibrosarcoma tissue.
Fixation: Formalin fixed paraffin sections
Positive Control: Human synovial, mammary, foreskin and thymic fibroblasts and in malignant fibrosarcoma tissue.
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


Not tested on frozen sections