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

Name: c-Myc (N-term)

Clone: Y69, Rabbit ant-Human

Supplier: Novus Biologicals

Catalog Number: NB110-57172

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.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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