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TintoFast EpCAM (Ber-EP4), MMab

Presentation:

Price: $347.00

SKU:	BSB 3678
Antibody type:	Mouse Monoclonal
Clone:	Ber-EP4
Isotype:	IgG1/K
Concentrate Dilution:	RTU
Localization:	Cytoplasmic

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Intended Use

For In Vitro Diagnostic Use

Summary and Explanation

Epithelial Cell Adhesion Molecule (EpCAM) or Epithelial Specific Antigen is a 40kD cell surface antigen that is broadly distributed in epithelial cells and displays a highly conserved expression in carcinomas. These glycoproteins are located on the cell membrane surface and in the cytoplasm of virtually all epithelial cells, with the exception of most squamous epithelia, hepatocytes, renal proximal tubular cells, gastric parietal cells and myoepithelial cells. However, focal positivity may be seen in the basal layer of squamous cell epithelium of endoderm (e.g., palatine tonsils) and mesoderm (e.g., uterine cervix).

EpCAM expression has been reported to be a possible marker of early malignancy, with expression being increased in tumor cells, and de novo expression being seen in dysplastic squamous epithelium. Epithelial specific antigen has been known to play an important role as a tumor-cell marker in lymph nodes from patients with esophageal carcinoma. EpCAM can be used to distinguish among Basal Cell, Basosquamous Carcinomas and Squamous Cell Carcinomas of the skin.

Presentation

Anti – EpCAM 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.