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Myosin VIIa Antibody Staining Protocol for Immunohistochemistry
Description: Myosin-VIIa is one of the unconventional members of the myosin molecular motor superfamily that move along filamentous actin. The protein is expressed primarily in the inner ear, retina and testis. Defects in the myosin-VIIa gene (MYO7A) are responsible for Usher syndrome, characterized by hearing impairment, lack of vestibular function and progressive retinal degeneration.MYO7A mutations are are responsible for deafness in shaker-1 mice and in other model systems. In the inner ear, myosin-VIIa is required for assembly of stereocilia, the sensory structures necessary for detecting sound signals.
Primary Antibody
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Name: Rabbit Anti-Myosin VIIa Antibody |
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Clone: Rabbit Polyclonal |
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Supplier: Proteus Biosciences |
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Catalog Number: 25-6790 |
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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. |
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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, Pig, Avian, Amphibian |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Inner ear, retina, 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: