Further Information - Leica BOND MSH6 Mode D'emploi

• The concentration of ProClin™ 950 is 0.35 %. It contains the active ingredient 2-methyl-4-isothiazolin-3-one, and may cause irritation
to the skin, eyes, mucous membranes and upper respiratory tract. Wear disposable gloves when handling reagents.
• To obtain a copy of the Material Safety Data Sheet contact your local distributor or regional office of Leica Biosystems, or alternatively,
visit the Leica Biosystems' Web site, www.LeicaBiosystems.com
• Specimens, before and after fixation, and all materials exposed to them, should be handled as if capable of transmitting infection and
disposed of with proper precautions
reagents or specimens. If reagents or specimens come in contact with sensitive areas, wash with copious amounts of water. Seek
medical advice.
• Consult Federal, State or local regulations for disposal of any potentially toxic components.
• Minimize microbial contamination of reagents or an increase in non-specific staining may occur.
• Retrieval, incubation times or temperatures other than those specified may give erroneous results. Any such change must be validated
by the user.
Instructions for Use
MSH6 (Mismatch Repair Protein) (EP49) primary antibody was developed for use on the automated BOND system (includes Leica
BOND-MAX system and Leica BOND-III system) in combination with BOND Polymer Refine Detection. The recommended staining
protocol for MSH6 (Mismatch Repair Protein) (EP49) primary antibody is IHC Protocol F. Heat-induced epitope retrieval is recommended
with BOND Epitope Retrieval Solution 2 for 20 minutes.
Results Expected
Normal Tissues
Clone EP49 detected the MSH6 protein in the nuclei of cells in a variety of tissues. Staining was observed in cerebrum, cerebellum,
adrenal gland, ovary, pancreas, parathyroid, pituitary, testis, thyroid, breast, spleen, tonsil, thymus, lung, esophagus, stomach, small
intestine, colon, salivary gland, kidney, prostate, endometrium, cervix, skin, peripheral nerves, mesothelium, eye, larynx and bladder.
(Total number of normal cases evaluated = 112).
Tumor Tissues
Clone EP49 stained 117/122 colorectal cancers, 1/1 adrenal gland tumor, 2/2 bladder tumors, 4/4 breast tumors, 2/2 bone tumors, 2/2
brain tumors, 2/2 tumors of the esophagus, 3/3 stomach tumors, 9/9 intestinal tract tumors, 1/1 kidney tumor, 3/3 liver tumors, 3/3 lung
tumors, 2/2 lymph node tumors, 3/3 head and neck tumors, 1/1 tongue tumor, 3/3 ovarian tumors, 1/1 pancreatic tumor, 2/2 prostate
tumors, 1/1 skin tumor, 1/1 testicular tumor, 4/4 thyroid tumors, 4/4 uterine tumors and 4/4 metastatic tumors. (Total number of abnormal
cases evaluated = 180).
PA0990 is recommended for the detection of the MSH6 protein in normal and neoplastic tissues, as an adjunct to conventional
histopathology using non-immunologic histochemical stains.
Product Specific Limitations
MSH6 (Mismatch Repair Protein) (EP49) has been optimized at Leica Biosystems for use with BOND Polymer Refine Detection and
BOND ancillary reagents. Users who deviate from recommended test procedures must accept responsibility for interpretation of patient
results under these circumstances. The protocol times may vary, due to variation in tissue fixation and the effectiveness of antigen
enhancement, and must be determined empirically. Negative reagent controls should be used when optimizing retrieval conditions and
protocol times.
Troubleshooting
Refer to reference 3 for remedial action.
Contact your local distributor or the regional office of Leica Biosystems to report unusual staining.

Further Information

Further information on immunostaining with BOND reagents, under the headings Principle of the Procedure, Materials Required,
Specimen Preparation, Quality Control, Assay Verification, Interpretation of Staining, Key to Symbols on Labels, and General Limitations
can be found in "Using BOND Reagents" in your BOND user documentation.
Bibliography
1. Clinical Laboratory Improvement Amendments of 1988, Final Rule 57 FR 7163 February 28, 1992.
2. Villanova PA. National Committee for Clinical Laboratory Standards (NCCLS). Protection of laboratory workers from infectious
diseases transmitted by blood and tissue; proposed guideline. 1991; 7(9). Order code M29-P.
3. Bancroft JD and Stevens A. Theory and Practice of Histological Techniques. 4th Edition. Churchill Livingstone, New York. 1996.
4. Offit K. MSH6 Mutations in Hereditary Nonpolyposis Colon Cancer: Another Slice of the Pie. Journal of Clinical Oncology. 2004;
22: 4449-4451.
5. Shia J, et al. Value of Immunohistochemical Detection of DNA Mismatch Repair Proteins in Predicting Germline Mutation in
Hereditary Colorectal Neoplasms. The American Journal of Surgical Pathology. 2005; 29: 96-104.
6. Shia J, et al. Immunohistochemistry as First-line Screening for Detecting Colorectal Cancer Patients at Risk for Hereditary
Nonpolyposis Colorectal Cancer Syndrome: A 2-antibody Panel May be as Predictive as a 4-antibody Panel. The American Journal
of Surgical Pathology. 2009; 33: 1639-1645.
7. Berends MJ, et al. Molecular and Clinical Characteristics of MSH6 Variants: An Analysis of 25 Index Carriers of a Germline Variant.
The American Journal of Human Genetics. 2002; 70: 26-37.
PA0990
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. Never pipette reagents by mouth and avoid contacting the skin and mucous membranes with
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