Data Sheet
Contents
1. Immunohistochemistry Assays Approved Guideline Second edition I LA28 A2 CLSI Wayne PA USA www clsi org 2011 Troubleshooting Follow the antibody specific protocol recommendations according to data sheet provided If atypical results occur contact Buiocare s Technical Support at 1 800 542 2002 EMERGO EUROPE Molenstraat 15 2513 BH The Hague The Netherlands Fax 925 603 8080 Tyrosinase ISO Concentrated and Prediluted Monoclonal Antibody prea na Control Number 901 155 032415 Precautions 1 This antibody contains less than 0 1 sodium azide Concentrations less than 0 1 are not reportable hazardous materials according to U S 29 CFR 1910 1200 OSHA Hazard communication and EC Directive 91 155 EC Sodium azide NaN used as a preservative is toxic if ingested Sodium azide may react with lead and copper plumbing to form highly explosive metal azides Upon disposal flush with large volumes of water to prevent azide build up in plumbing Center for Disease Control 1976 National Institute of Occupational Safety and Health 1976 5 2 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 Never pipette reagents by mouth and avoid contacting the skin and mucous membranes with reagents and specimens If reagents or specimens come in contact with sensitive areas wash with copious amounts of water 6 3 Mic2. System User Manual for specific instructions on its use Protocol parameters in the ONCORE Automated Slide Stainer Protocol Editor should be programmed as follows Protocol Name Tyrosinase Protocol Template Description Ms HRP Template 1 Dewaxing DS Option DS2 Antigen Retrieval AR Option AR2 low pH 101 C Reagent Name Time Temp Tyrosinase 30 min 25 C Limitations The optimum antibody dilution and protocols for a specific application can vary These include but are not limited to fixation heat retrieval method incubation times tissue section thickness and detection kit used Due to the superior sensitivity of these unique reagents the recommended incubation times and titers listed are not applicable to other detection systems as results may vary The data sheet recommendations and protocol are based on exclusive use of Biocare products Ultimately it is the responsibility of the investigator to determine optimal conditions The clinical interpretation of any positive or negative staining should be evaluated within the context of clinical presentation morphology and other histopathological criteria by a qualified pathologist The clinical interpretation of any positive or negative staining should be complemented by morphological studies using proper positive and negative internal and external controls as well as other diagnostic tests Quality Control Refer to CLSI Quality Standards for Design and Implementation of
3. tested In conclusion tyrosinase is shown to be a superior melanoma marker when compared to HMB 45 Principle of Procedure Antigen detection in tissues and cells is a multi step immunohistochemical process The initial step binds the primary antibody to its specific epitope A secondary antibody may be applied to bind the primary antibody followed by an enzyme labeled polymer or an enzyme labeled polymer may be applied directly to bind the primary antibody The detection of the bound primary antibody is evidenced by an enzyme mediated colorimetric reaction Source Mouse monoclonal Species Reactivity Human others not tested Clone T311 Isotype IgG2a Total Protein Concentration 10 mg ml Call for lot specific Ig concentration Epitope Antigen Tyrosinase Cellular Localization Cytoplasm Positive Control Melanoma Known Applications Immunohistochemistry formalin fixed paraffin embedded tissues Supplied As Buffer with protein carrier and preservative Storage and Stability Store at 2 C to 8 C Do not use after expiration date printed on vial If reagents are stored under conditions other than those specified in the package insert they must be verified by the user Diluted reagents should be used promptly any remaining reagent should be stored at 2 C to 8 C Protocol Recommendations manual use Peroxide Block Block for 5 minutes with Biocare s Peroxidazed 1 Pretreatment Solution recommended Reveal Pretreatme
4. Tyrosinase Concentrated and Prediluted Monoclonal Antibody ISO 9001813485 CERTIFIED Control Number 901 155 032415 Catalog Number CM 155 A B C PM 155 AA Description 0 1 0 5 1 0 ml concentrated 6 0 ml prediluted Dilution 1 50 1 200 Ready to use Diluent Van Gogh Yellow N A Intended Use For In Vitro Diagnostic Use Tyrosinase T311 is a mouse monoclonal antibody that is intended for laboratory use in the qualitative identification of tyrosinase protein by immunohistochemistry IHC in formalin fixed paraffin embedded FFPE human tissues The clinical interpretation of any staining or its absence should be complemented by morphological studies using proper controls and should be evaluated within the context of the patient s clinical history and other diagnostic tests by a qualified pathologist Summary and Explanation Tyrosine is a key enzyme involved in the initial stages of melanin biosynthesis Studies have shown tyrosinase to be a more sensitive marker when compared to HMB 45 and MART 1 It has also been shown to label a higher percentage of desmoplastic melanomas than HMB 45 However both tyrosinase and MART 1 negative staining was seen in those variants without an epidermal component Unlike HMB 45 tyrosinase or MART 1 does not discriminate between activated or resting melanocytes Other studies have shown tyrosinase to be a very specific marker for melanomas and did not cross react with any tumors or normal tissues
5. nt Protocol Heat Retrieval Method Retrieve sections under pressure using Biocare s Decloaking Chamber followed by a wash in distilled water alternatively steam tissue sections for 45 60 minutes Allow solution to cool for 10 minutes then wash in distilled water taal Biocare Medical 4040 Pike Lane Concord CA 94520 USA Tel 800 799 9499 l CE Page of 2 www biocare net OAI 155 T60 60 tests prediluted Ready to use N A Protocol Recommendations manual use Cont d Protein Block Optional Incubate for 5 10 minutes at RT with Biocare s Background Punisher Primary Antibody Incubate for 15 30 minutes at RT Probe Incubate for 10 minutes at RT with a secondary probe Polymer Incubate for 10 minutes at RT with a tertiary polymer Chromogen Incubate for 5 minutes at RT with Biocare s DAB OR Incubate for 5 7 minutes at RT with Biocare s Warp Red Counterstain Counterstain with hematoxylin Rinse with deionized water Apply Tacha s Bluing Solution for 1 minute Rinse with deionized water Technical Note This antibody has been standardized with Biocare s MACH 4 detection system It can also be used on an automated staining system and with other Biocare polymer detection kits Use TBS buffer for washing steps Protocol Recommendations OQNCORE Automated Slide Staining System OAII55 is intended for use with the ONCORE Automated Slide Staining System Refer to the ONCORE Automated Slide Staining
6. robial contamination of reagents may result in an increase in nonspecific staining 4 Incubation times or temperatures other than those specified may give erroneous results The user must validate any such change 5 Do not use reagent after the expiration date printed on the vial 6 The SDS is available upon request and is located at http biocare net support References 1 Orchard GE Comparison of immunohistochemical labelling of melanocyte differentiation antibodies melan A tyrosinase and HMB 45 with NKIC3 and S100 protein in the evaluation of benign naevi and malignant melanoma Histochem J 2000 Aug 32 8 475 81 2 Jungbluth AA et al T311 an anti tyrosinase monoclonal antibody for the detection of melanocytic lesions in paraffin embedded tissues Pathol Res Pract 2000 196 4 235 42 3 Kaufmann O et al Tyrosinase melan A and KBA62 as markers for the immunohistochemical identification of metastatic amelanotic melanomas on paraffin sections Mod Pathol 1998 Aug 11 8 740 6 4 Hofbauer GF et al Tyrosinase immunoreactivity in formalin fixed paraffin embedded primary and metastatic melanoma frequency and distribution J Cutan Pathol 1998 Apr 25 4 204 9 5 Center for Disease Control Manual Guide Safety Management NO CDC 22 Atlanta GA April 30 1976 Decontamination of Laboratory Sink Drains to Remove Azide Salts 6 Clinical and Laboratory Standards Institute CLSI Protection of Laboratory Workers from Occ
7. upationally Acquired Infections Approved Guideline Fourth Edition CLSI document M29 A4 Wayne PA 2014 pool Biocare Medical EMERGO EUROPE 4040 Pike Lane CE Molenstraat 15 Concord CA 94520 2513 BH The Hague Page 2 of 2 USA The Netherlands Tel 800 799 9499 www biocare net Fax 925 603 8080
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