Myosin VIIa Antibody Staining Protocol for Immunohistochemistry

 

Description: Myosin VIIa is a member of the myosin superfamily of actin-based motor proteins. Defects in the myosin VIIa gene are responsible for hearing impairment in shaker-1 (sh1) mice and causes Usher syndrome IB in humans. Usher syndrome associates congenital deafness, vestibular dysfunction, and retinitis pigmentosa and is the most common form of combined deafness and blindness. Structural features of myosin VIIa protein include an ATP binding N-terminal motor domain, a central region which possess five light-chain binding (IQ) motifs, and a C-terminal domain with three myosin tail homology (MyTH4) and talin-like homology regions. This antibody Detects Myosin VIIa from mouse tissues as well as recombinant. By Western blot, this antibody detects an ~220 kDa protein representing myosin VIIa from mouse testes preparations. This antibody detects recombinant mouse myosin VIIa overexpressed in Sf9 insect cell lysate.

 

Primary Antibody

Name: Rabbit Anti-Myosin VIIa Antibody

Clone: Rabbit Polyclonal

Supplier: Abcam

Catalog Number: ab3481

Dilution: 1:500 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, Dog, Pig
Fixation: Formalin fixed paraffin sections
Positive Control: Mouse inner ear, testis
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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