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Androgen Receptor (AR) Antibody Staining Protocol for Immunohistochemistry
Description: The androgen receptor (AR) is a NR3 Steroid Receptor located on the X chromosome. AR is a phosphoprotein and acts as a steroid hormone-activated transcription factor for androgen-responsive genes. Mutations in the AR gene are associated with androgen insensitivity syndrome (CAIS) or testicular feminization syndrome, Reifenstein syndrome, and Kennedy spinal and bulbar muscular atrophy. They lead to symptoms such as low virilization, reduced sperm production, testicular atrophy, and infertility. Immunohistochemistry: LS-B3326 was validated for use in immunohistochemistry on a panel of 21 formalin-fixed, paraffin-embedded (FFPE) human tissues after heat induced antigen retrieval.
Primary Antibody
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Name: Rabbit Anti-AR Antibody |
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Clone: Rabbit Polyclonal |
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Supplier: LifeSpan BioSciences |
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Catalog Number: LS-B3326 |
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Dilution: 1:200 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 Chromogen Substrate |
| 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, mouse, rat |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Human prostate |
| 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: