Data Sheet
Contents
1. CD8 Concentrated and Prediluted Monoclonal Antibody ISO 9001813485 CERTIFIED Control Number 901 154 032415 Catalog Number CM 154 A C PM 154 AA Description 0 1 1 0 ml concentrated 6 0 ml prediluted Dilution 1 75 1 150 Ready to use Diluent Van Gogh Yellow N A Intended Use CD8 BC 1A5 is a mouse monoclonal antibody that is intended for laboratory use in the qualitative identification of CD8 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 CD8 antibody reacts with a 32 kDa protein equivalent to the CD8 molecule CD8 stains cells with cytotoxic activity including cytotoxic suppressor T cells and a subset of natural killer cells CD4 and CD8 positive and negative staining are indicative of T cell neoplasms CD4 and CD8 are also used to differentiate between mycosis fungoides and cutaneous inflammatory processes CD8 is also used in panels with CD3 CD4 CD57 and TIA 1 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 prima2. 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 3 EMERGO EUROPE Molenstraat 15 2513 BH The Hague The Netherlands Fax 925 603 8080 CD8 Concentrated and Prediluted Monoclonal Antibody Control Number 901 154 032415 Precautions Cont d Troubleshooting 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 4 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 1 800 542 2002 References 1 Barth TF et al Primary gas
3. however users must validate incubation times and protocols for their specific application 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 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 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
4. ry 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 BC 1A5 Isotype IgGl Total Protein Concentration 10 mg ml Call for lot specific Ig concentration Epitope Antigen CD8 Cellular Localization Cell membrane Positive Control Tonsil or T cell lymphoma 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 Diva 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 10 minutes then wash in distilled water Protein Block Optional Incubate for 5 10 minu
5. tes 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 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 OAI 154 T60 60 tests prediluted Ready to use N A Protocol Recommendations 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 Protocol Recommendations ONCORE Automated Slide Staining System OAI154 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 Protocol parameters in the ONCORE Automated Slide Stainer Protocol Editor should be programmed as follows Protocol Name CD8 Protocol Template Description Ms HRP Template 1 Dewaxing DS Option DS2 Antigen Retrieval AR Option AR1 high pH 103 C Reagent Name Time Temp CD8 30 min 25 C Technical Note This antibody has been optimized for use with Biocare s MACH 4 Universal HRP Polymer Detection and ONCORE HRP Detection Other Biocare polymer detection kits may be used
6. tric apoptosis rich T cell lymphoma co expressing CD4 CD8 and cytotoxic molecules Virchows Arch 2000 Apr 436 4 357 64 2 Williamson SL et al New monoclonal antibodies to the T cell antigens CD4 and CD8 Production and characterization in formalin fixed paraffin embedded tissue Am J Pathol 1998 Jun 152 6 1421 6 3 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 4 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 ISO 9001813485 CERTIFIED Follow the antibody specific protocol recommendations according to data sheet provided If atypical results occur contact Buiocare s Technical Support at EMERGO EUROPE Molenstraat 15 2513 BH The Hague The Netherlands
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