Intended Use
For In Vitro Diagnostic Use
Summary and Explanation
The androgen receptor (AR) is a type of nuclear receptor which is activated by binding of either of the androgenic hormones testosterone or dihydrotestosterone. The main function of the androgen receptor is as a DNA-binding transcription factor which regulates gene expression. However, the androgen receptor has additional functions independent of DNA binding. The AR signaling pathway plays a key role in development and function of male reproductive organs, including the prostate and epididymis. AR also plays a role in nonreproductive organs, such as muscle, hair follicles, and the brain.
TintoFast Androgen Receptor antibody reacts with the androgen receptor and also with the newly-described A form of the receptor. This antibody does not cross-react with estrogen, progesterone or glucocorticoid receptors. Abnormalities in the AR-signaling pathway have been linked to a number of diseases, including Prostate Cancer, Kennedy’s Disease and male infertility.
Presentation
TintoFast Androgen Receptor is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.
Mohs IHC Procedure
Specimen Preparation of Mohs Frozen Tissues
- Embed the specimen in OCT inside a cryostat.
- Cut sections at 4-5 µm and mount on a positively charged glass slide such as the Bio SB Hydrophilic Plus Slides (BSB 7028) or in the lower third of the TintoDetector Cap Gap slides (BSB 7006).
- Air dry the slide at room temperature for 2 minutes and then incubate the slide at 60 °C for 3 minutes in an incubator or dry bath.
- Fix in 100% acetone for 2 minutes at room temperature.
Pretreatment of Mohs Frozen Tissues
- Preheat the TintoDetector Incubator to 110°C.
- Place TintoDetector Cap Gap slides (BSB 7006) face to face and insert them into the TintoDetector Slide Holder (BSB 7003).
- Submerge slides in ImmunoDNA Retriever with EDTA to draw up enough solution by capillary action to cover the tissues.
- Heat the slides in a preheated TintoDetector Incubator for 3 minutes.
- Transfer slides to room temperature and cool off for 1 min.
Mohs IHC Detection
- After HIER, transfer slides to ImmunoDNA washer and let it stand for 1-2 minutes.
- For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.
- Wash slides with ImmunoDNA washer or DI water.
- Continue IHC detection protocol. Wash slides between each step with ImmunoDNA washer solution.
Abbreviated Mohs PolyDetector Plus DAB HRP Brown or HRP Green Immunohistochemical Protocol
- Incubate with Primary Antibody for 5 min
- Wash with Buffer (TBST or PBST)
- Incubate with M/R link for 4 min
- Wash with Buffer (TBST or PBST)
- HRP Label for 4 min.
- Wash with Buffer (TBST or PBST)
- Prepare
- DAB Brown (1 drop of DAB Chromogen in 1ml of DAB Buffer; mix well)
- or HRP Green (1 drop of HRP Green Chromogen in 1 ml of HRP Green Buffer; mix well)
- Incubate with DAB or HRP Green for 1-2 min
- Wash with Buffer (TBST or PBST)
- Counterstain with Hematoxylin or Nuclear Fast Red for 30 seconds
- Wash with Buffer (TBST or PBST)
- Mount with AquaMounter or dehydrate the tissue with Fast ChromoProtector then mount with PermaMounter
Step
|
Mohs PolyDetector Plus HRP Green or DAB 20 Minute Protocol |
| HIER |
3 min. |
| Primary Antibody |
5 min. |
| 1st Step Detection |
4 min. |
| 2nd Step Detection |
4 min. |
| Substrate-Chromogen |
1-2 min. |
| Counterstain / Coverslip |
Varies |
This protocol can also be used with FFPE tissues retrieved with Citrate or EDTA.
