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Progesterone Receptor (PR) Antibody Staining Protocol for Immunohistochemistry
Description: The role of steroid hormone receptors in breast cancer is well known. The absence of ER and PR predicts early recurrence and poor survival of breast cancer patients. Also, the presence of ER and PR in tumors predicts the potential for benefit from tamoxifen and other endocrine-related therapies. Measurement of ER and PR can be determined semi-quantitatively using IHC or quantitatively using DCC or EIA. Correlation between the semi-quantitative and quantitative evaluations of PR have ranged from 73 to 91% depending on the laboratory and antibody used.
Primary Antibody
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Name: Progesterone Receptor (PR) Antibody |
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Clone: Rabbit anti-Human |
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Supplier: Dako |
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Catalog Number: A0098 |
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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: 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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Nuclear |
| Images: Search image |
Additional Information:
| Species Reactivity: Human |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections (no antigen retrieval needed) |
| Positive Control: Uterine gland cells, breast, prostate, endometrium |
| 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: