Desmin Antibody Staining Protocol for Immunohistochemistry

 

Description: Desmin is a 53 kDa intermediate filament protein present in smooth muscle cells, striated muscle cells and myocardium. In skeletal and cardiac muscle cells, desmin filaments are localized in the Z-bands.

 

Primary Antibody

Name: Desmin Antibody

Clone: DE-R-11, Mouse anti-Porcine

Supplier: Dako

Catalog Number: M0724

Dilution: 1:30 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: Cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Skeletal and cardiac muscle
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Skeletal and cardiac muscle cells, smooth muscle cells of blood vessels, viscera, the uterus as well as in Purkinje cells, many mesangial cells of the kidney glomeruli, leiomyoma and rhabdomyomas.
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:

No need to perform antigen retrieval on frozen sections

 

References: