IHC of Cytokeratin MNF116 on a Frozen NBF 10%-Fixed Basal Cell Carcinoma Tissue

Specification Sheets
Safety Data Sheet

SDS

Intended Use For Mohs In Vitro Diagnostic Use
Summary and Explanation Cytokeratin MNF116 is a broad-spectrum anti-cytokeratin reacting with intermediate and low-molecular-weight keratins, ranging from 40 through 58 kD, corresponding to cytokeratin 5, 6, 8, 17 and 19. It shows a broad pattern of reactivity with human epithelial tissues from simple glandular epithelial to stratified squamous epithelia, like epidermis, mammary gland ducts, and tracheal epithelium.

Cytokeratin MNF116 is a useful aid for the classification of neoplasms of epithelial origin including Squamous Cell Carcinoma, Small Cell Carcinoma, Sarcomatoid Carcinoma, Spindle Cell Carcinoma, Epithelioid and Spindle Cell component of Malignant Mesothelioma and Adenocarcinoma. A wide range of soft tissue tumors are also positive with cytokeratin MNF116: monophasic and biphasic Synovial Sarcoma, vascular neoplasms including Epithelioid Hemangioendothelioma, Epithelioid Angiosarcoma, Epithelioid Sarcoma. Desmoplastic Small Round Cell Tumors require cytokeratin positivity for diagnosis. Smooth muscle tumors and Plasmacytoma may demonstrate aberrant expression of cytokeratin MNF116.

Antibody Type Mouse Monoclonal Clone MNF116
Isotype IgG1 Reactivity Paraffin, Frozen
Localization Cytoplasmic Control SCC, BCC
Presentation Anti – Cytokeratin MNF116 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.
Availability
Catalog No. Antibody Type Dilution Volume/QTY
BSB 3672 TintoFast Prediluted Ready-To-Use 3.0 ml
BSB 3673 TintoFast Prediluted Ready-To-Use 7.0 ml
BSB 3674 TintoFast Prediluted Ready-To-Use 15.0 ml

Mohs IHC Procedure

Specimen Preparation of Mohs Frozen Tissues

  1. Embed the specimen in OCT inside a cryostat.
  2. 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).
  3. 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.
  4. Fix in 100% acetone or 10% NBF for 2 minutes at room temperature. Using NBF 10% produces better morphology.
  5. Rinse with distilled water and air dry the slides for another 2 minutes at room temperature.

IHC Detection Procedure

  1. Transfer slides to ImmunoDNA washer, TBST or PBST buffer.
  2. For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.
  3. Wash slides with ImmunoDNA washer, TBST, PBST or DI water.
  4. Continue IHC detection protocol. Wash slides between each step with ImmunoDNA washer, Tris or PBS Buffer solution.

Abbreviated Mohs Immunohistochemical Protocol

  1. Do HIER with Citrate in Pressure Cooker (100 – 121 °C ) for 1 min. Cool off
  2. Wash with Buffer (TBST or PBST)
  3. Do PIER, proteolytic Digestion, with Mohs ImmunoDigestor for 30 sec.
  4. Wash with Buffer (TBST or PBST)
  5. Incubate slides with Peroxidase Blocker for 30 seconds
  6. Wash with Buffer (TBST or PBST)
  7. Incubate CK MNF116 for 4 min
  8. Wash with Buffer (TBST or PBST)
  9. Incubate with HRP Label for 3 min.
  10. Wash with Buffer (TBST or PBST)
  11. Prepare
    • DAB  Brown (1 drop of DAB chromogen in 1 ml of DAB Buffer; mix well)
    • or HRP Green (1 drop of HRP Green chromogen in 1 ml of HRP Green Buffer, mix well)
  1. Incubate with DAB or HRP Green for 2 min
  2. Wash with Buffer (TBST or PBST)
  3. Counterstain with Hematoxylin or Nuclear Fast Red for 30 seconds
  4. Wash with Buffer (TBST or PBST)
  5. Mount with AquaMounter or PermaMounter

For best results with the IHC of CK MNF116, we recommend using Mohs Frozen sections fixed NBF 10% for 2 min., then HIER with Citrate at 100 – 110 °C  for 2 min., plus PIER with Mohs ImmunoDigestor (BSB 0324 – 0326 )at room temperature for 30 sec.

Step Mohs PolyDetector HRP Green 5 min Protocol Mohs PolyDetector HRP Green 10 min Protocol
Peroxidase Blocker 0.5 min. 0.5 min.
Primary Antibody 2 min 4 min.
1st Step Detection 1 min 3 min.
Substrate-Chromogen 1 min 2 min.
Counterstain / Coverslip 0.5 min 0.5 min.

This protocol can also be used with FFPE Tissues retrieved with Citrate or EDTA.

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