Muscle Specific Actin (MSA)
Contents
1. 10 minutes then wash in distilled water Protein Block Incubate for 20 minutes at RT with Biocare s Background Punisher Primary Antibody Incubate for 30 minutes at RT Probe Incubate for 10 minutes at RT with a secondary probe Polymer Incubate for 10 20 minutes at RT with a tertiary polymer taal Biocare Medical 4040 Pike Lane Concord CA 94520 USA Tel 800 799 9499 l CE Page of 2 www biocare net IP 079 G10 10 ml prediluted OAI 079 T60 60 tests prediluted Ready to use N A Ready to use N A Protocol Recommendations intelliPATH and manual use Cont d 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 intelliPATH Automated Slide Stainer IP079 is intended for use on the intelliPATH Automated Slide Stainer Refer to the intelliPATH Automated Slide Stainer manual for specific instructions on its use When using the intelliPATH peroxide block with intelliPATH Peroxidase Blocking Reagent IPB5000 may be performed following heat retrieval Protocol Recommendations ONCORE Automated Slide Staining System OAIO079 is intended for use with the ONCORE Automated Slide Staining System Refer to the ONCORE Automated Slide Staining System User Manual for specific instructions on its use Pr2. Remove Azide Salts 6 Clinical and Laboratory Standards Institute CLSI Protection of Laboratory Workers from Occupationally Acquired Infections Approved Guideline Fourth Edition CLSI document M29 A4 Wayne PA 2014 aol Biocare Medical 4040 Pike Lane CE Concord CA 94520 Page 2 of 2 USA Tel 800 799 9499 www biocare net Fax 925 603 8080 EMERGO EUROPE Molenstraat 15 2513 BH The Hague The Netherlands
3. Muscle Specific Actin MSA Concentrated and Prediluted Monoclonal Antibody ISO 9001813485 CERTIFIED Control Number 901 079 102015 Catalog Number CM 079 A B PM 079 AA Description 0 1 0 5 ml concentrated 6 0 ml prediluted Dilution 1 50 1 100 Ready to use Diluent Da Vinci Green N A Intended Use For In Vitro Diagnostic Use Muscle Specific Actin MSA HHF35 is a mouse monoclonal antibody that is intended for laboratory use in the qualitative identification of muscle actin 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 The HHF 35 antibody recognizes muscle specific alpha and gamma actin isomers It does not react with non muscle actin It recognizes the alpha actin from cardiac skeletal and smooth muscle sources It does not react with beta or nonsmooth muscle gamma actin isomers Studies have shown the antibody labels leiomyoma leiomyosarcoma and rhabdomyosarcoma It does not label melanoma or lymphoma 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 antibo
4. dy 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 HHF35 Isotype IgG1 kappa Total Protein Concentration 10 mg ml Call for lot specific Ig concentration Epitope Antigen Muscle actin Cellular Localization Cytoplasmic Positive Tissue Control Leiomyoma leiomyosarcoma muscle or prostate 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 intelliPA TH and manual use Peroxide Block Block for 5 minutes with Biocare s Peroxidazed 1 Pretreatment Solution recommended Reveal Pretreatment 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
5. otocol parameters in the ONCORE Automated Slide Stainer Protocol Editor should be programmed as follows Protocol Name MSA Protocol Template Description Ms HRP Template 1 Dewaxing DS Option DS2 Antigen Retrieval AR Option AR2 low pH 25 C Reagent Name Time Temp MSA 30 min 25 C Technical Note This antibody has been optimized for use with Biocare s MACH 4 Universal HRP Polymer Detection intelliPATH Universal HRP Detection Kit and ONCORE HRP Detection Use TBS for washing steps 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 protocols 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 e
6. 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 Microbial 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 References 1 Rangdaeng S Truong LD Comparative immunohistochemical staining for desmin and muscle specific actin A study of 576 cases Am J Clin Pathol 1991 Jul 96 1 32 45 2 Miettinen M Antibody specific to muscle actins in the diagnosis and classification of soft tissue tumors Am J Pathol 1988 Jan 130 1 205 15 3 Tsukada T et al HHF35 a muscle actin specific monoclonal antibody II Reactivity in normal reactive and neoplastic human tissues J Pathol 1987 May 127 2 389 402 4 Norton AJ Thomas JA Isaacson PG Cytokeratin specific monoclonal antibodies are reactive with tumours of smooth muscle derivation An immunocytochemical and biochemical study using antibodies to intermediate filament cytoskeletal proteins Histopathology 1987 May 11 5 487 99 5 Center for Disease Control Manual Guide Safety Management NO CDC 22 Atlanta GA April 30 1976 Decontamination of Laboratory Sink Drains to
7. xternal controls as well as other diagnostic tests Quality Control Refer to CLSI Quality Standards for Design and Implementation of Immunohistochemistry Assays Approved Guideline Second edition I LA28 A2 CLSI Wayne PA USA www clsi org 2011 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 EMERGO EUROPE Molenstraat 15 2513 BH The Hague The Netherlands Fax 925 603 8080 Muscle Specific Actin MSA ISO Concentrated and Prediluted Monoclonal Antibody ee Control Number 901 079 102015 Troubleshooting Precautions Cont d Follow the antibody specific protocol recommendations according to data sheet volumes of water to prevent azide build up in plumbing Center for Disease Control provided If atypical results occur contact Biocare s Technical Support at 1976 National Institute of Occupational Safety and Health 1976 5 1 800 542 2002 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
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