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

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

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

Notes:

Not tested on frozen sections

 

References: