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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
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Name: Rabbit Anti-Myosin VIIa Antibody |
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Clone: Rabbit Polyclonal |
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Supplier: Abcam |
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Catalog Number: ab3481 |
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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. |
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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 |
References: