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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
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Name: Aurora-A/STK15 Antibody |
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Clone: Rabbit polyclonal |
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Supplier: TransGenic |
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Catalog Number: KR-051 |
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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: 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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
Notes:
There is no need to perform antigen retrieval on acetone fixed frozen sections |
References: