Actin (Muscle) Antibody Staining Protocol for Immunohistochemistry

 

Description: This antibody reacts with the muscle actin on formalin fixed paraffin embedded sections or frozen sections.

 

Primary Antibody

Name: Mouse Anti-Human Actin, Muscle

Clone: HHF35

Supplier: Dako

Catalog Number: M0635

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 min/room temperature


Antigen Retrieval
Device: Antigen retrieval is not 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: Membrane/Cytoplasmic
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin-fixed paraffin sections or acetone fixed frozen sections
Positive Control: Skeletal muscle, tongue, leg muscle
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

Notes:

There is no need to perform antigen retrieval on acetone fixed frozen sections

 

References: