Myosin VIIa Antibody Staining Protocol for Immunohistochemistry

 

Description: Myosins are molecular motors that move along filamentous Actin and influence cellular movements such as phagocytosis. There are seven classes of myosins in vertebrates, including Myosin II, and six unconventional Myosin classes, designated I, V, VI, VII, IX and X. Myosin VIIa is a plus end-directed motor that influences cilia formation and cell adhesion. Mutations in the human Myosin VIIa gene correlate with Usher syndrome, a disease characterized by congenital sensorineural deafness, vestibular dysfunction and retinitis pigmentosa.

 

Primary Antibody

Name: Mouse Anti-Myosin VIIa Antibody

Clone: Mouse Monoclonal (C-5)

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-74516

Dilution: 1:100 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: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 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:
Species Reactivity: Human, Mouse, Rat
Fixation: Formalin fixed paraffin sections
Positive Control: Mouse inner ear, human pancreas
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

 

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