Cdc2 Antibody Staining Protocol for Immunohistochemistry


Description: During the eukaryotic cell cycle, activation of Cdc2 (Cdk1) kinase regulates the transition into M-phase. Cdc2 forms a complex with cyclin B, and activation of this complex is controlled by dephosphorylation of Cdc2 at tyrosine 15 and threonine 14. Phosphorylation at Tyr15 and is carried out by Wee1 and Myt1 protein kinases, while Tyr15 dephosphorylation is carried out by the cdc25 phosphatase.

Primary Antibody

Name: Cdc2 (Y15) (Non-phospho-specific) Antibody

Clone: Rabbit monoclonal

Supplier: Epitomics

Catalog Number: 1161-1

Dilution: 1:50 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: 95-100 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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasmic/nuclear
Images: Search image

Additional Information:
Tissue Type: Tonsil, lymphoma, colorectal cancer
Fixation: Formalin fixed paraffin sections
Positive Control: Tonsil, lymphoma, colorectal cancer
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



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