Aurora A/STK15 Antibody Staining Protocol for Immunohistochemistry


Description: Aurora-A (Aurora-A/STK-15/Aik) encoding a centrosome-associated kinase, is amplified and over-expressed in many cancer cell types, and is involved in the induction of centrosome duplication-distribution abnormalities and aneuploidy in mammalian cells. So that, Aurora-A is a potential oncogene. Aurora-A have been identified three types (Aurora-A, B, C) which has high homology domain in C terminal region. Aurora family is indispendsable kinase that regulate the structure and function of centrosomes and spindle. Aurora-A could be one of targeting molecule of anti-cancer therapy. This antibody was established from the purified serum of the immunized rabbit with N terminal region of Aurora-A and did not show any cross-reactivity to Aurora-B and Aurora-C. This antibody is useful for western blotting and immunohistochemistry.


Primary Antibody

Name: Aurora-A/STK15 Antibody

Clone: Rabbit polyclonal

Supplier: TransGenic

Catalog Number: KR-051

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.

Incubation Time/Temp: 60min/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) or Proteinase K Solution (Cat# IW-1101)
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/membrane
Images: Search image

Additional Information:
Tissue Type: MCF-7 cells
Fixation: Formalin-fixed paraffin sections, or acetone fixed frozen sections
Positive Control: MCF-7 cells
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


There is no need to perform antigen retrieval on acetone fixed frozen sections