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

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: Gill's Hematoxylin or Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

Staining Pattern: Membrane/Cytoplasmic

Images: Search image

Additional Information:

Tissue Type: Leg muscle

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: