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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
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Name: Mouse Anti-Myosin VIIa Antibody |
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Clone: Mouse Monoclonal (C-5) |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-74516 |
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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 |
| 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 |
References: