Intended Use
For In Vitro Diagnostic Use
Summary and Explanation
Adipose differentiation-related protein, also known as Perilipin 2 (PLIN2), ADRP or Adipophilin, is a protein which in humans is encoded by the ADFP gene. Adipophilin is associated with the globule surface membrane material. This protein is a major constituent of the globule surface. Increase in mRNA levels is one of the earliest indications of adipocyte differentiation. Adipophilin is present in a wide range of cultured cell lines, including fibroblasts, endothelial, and epithelial cells. In tissues, however, expression of Adipophilin is restricted to certain cell types, such as Lactating Mammary epithelial cells, Adrenal Cortex cells, Sertoli and Leydig cells of the male reproductive system, and Steatosis or fatty change hepatocytes in alcoholic liver Cirrhosis.
Adipophilin antibody expression in various Sebaceous lesions and other Cutaneous Tumors with a clear cell histology that may mimic sebaceous differentiation. Adipophilin can be valuable in an immunohistochemical panel when evaluating cutaneous lesions with clear cell histology as it identifies intracytoplasmic lipid vesicles in Sebaceous and Xanthomatous lesions. In periocular lesions, it is effective in helping to exclude Basal Cell Carcinoma and Squamous Cell Carcinoma when sebaceous carcinoma is under consideration. Adipophilin expression is not as useful for the differential diagnosis that includes Metastatic Renal Cell Carcinoma, a rare but important, diagnostic differential. The pattern of adipophilin reactivity is important to observe as membranous vesicular staining is suggestive of intracellular lipids whereas granular cytoplasmic reactivity is not. Adipophilin antibody is suitable for immunostaining and is helpful in the identification of intracytoplasmic lipids, as seen in Sebaceous lesions. It is especially helpful in identifying intracytoplasmic lipid vesicles in poorly differentiated Sebaceous Carcinomas in challenging cases such as small periocular biopsy specimens.
Presentation
Adipophilin 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.
