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c-Myc Antibody Staining Protocol for Immunohistochemistry
Description: Oncogene-encoded proteins c-Myc, N-Myc, and L-Myc function in cell proliferation, differentiation and neoplastic disease. Amplification of the c-Myc gene has been found in several types of human tumors, the N-Myc gene in neuroblastomas, and the L-Myc gene in human small cell lung carcinomas. c-Myc protein is a transcription factor localized to the nucleus of the cell. It seems to be involved in activating the transcription of growth-related genes. c-Myc binds to DNA during transcription as a heterodimeric complex with Max. c-Myc is phosphorylated in vitro by p44/42 MAP kinase at Ser62 and in vivo at both Thr58 and Ser62. Mutation of Thr58 and Ser62 to Ala inhibits the ability of c-Myc to activate transcription.
Primary Antibody
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Name: c-Myc (N-term) |
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Clone: Y69, Rabbit ant-Human |
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Supplier: Novus Biologicals |
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Catalog Number: NB110-57172 |
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Dilution: 1:50 - 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 ºC - 100 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minuters |
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: Nuclear |
| Images: Search image |
Additional Information:
| Species Reactivity: Human. Does not react in mouse or rat. |
| Fixation: Formalin fixed paraffin sections. |
| Positive Control: Human tumors - neuroblastomas, colon, small cell lung carcinomas. |
| 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: