Data Sheet
Contents
1. 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 Netherlands2. Tacha s Bluing Solution for 1 minute Rinse with deionized water intelliPATH Automated Slide Stainer IP162 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 pretreatment Protocol Recommendations ONCORE Automated Slide Staining System OAI162 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 Pan CK Plus Protocol Template Description Ms HRP Template 1 Dewaxing DS Option DS Buffer Antigen Retrieval AR Option AR2 low pH 90 C Reagent Name Time Temp Pan CK Plus 30 min 25 C Technical Note 1 With cytokeratin markers heat retrieval may provide a higher sensitivity assay whereas enzyme digestion may produce greater specificity Users should validate the pretreatment method for their specific application 2 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 Other Biocare polymer detection kits may be used however users must validate i
3. Pan Cytokeratin Plus AE1 AE3 8 18 Concentrated and Prediluted Cocktail Antibody ISO 9001813485 CERTIFIED Control Number 901 162 032415 Catalog Number CM 162 A B C PM 162 AA H Description 0 1 0 5 1 0 ml concentrated 6 0 25 ml prediluted Dilution 1 50 1 100 Ready to use Diluent Da Vinci Green N A Intended Use For In Vitro Diagnostic Use Pan Cytokeratin Plus AE1 AE3 8 18 is a mouse monoclonal antibody cocktail that is intended for laboratory use in the qualitative identification of a broad spectrum of acidic and basic cytokeratin proteins 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 AEI1 AE3 recognizes acidic and basic subfamilies of cytokeratins The cocktail of these two antibodies can be used to detect most human epithelia The acidic cytokeratins have molecular weights of 56 5 55 51 50 50 48 46 45 and 40 kDa The basic cytokeratins have molecular weights of 65 67 64 59 58 56 and 52 kDa Clone 5D3 recognizes cytokeratin CK 8 and 18 intermediate filament proteins These are 52 5 kDa and 45 kDa respectively In normal tissues 5D3 recognizes all simple and glandular epithelium In
4. ibody Control Number 901 162 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 Troubleshooting a a aes 1 800 542 2002 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 6 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 into contact with sensitive areas wash with copious amounts of water 7 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 Seidman JD Abbondanzo SL Bratthaue
5. ncubation 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 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 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 EMERGO EUROPE Molenstraat 15 2513 BH The Hague The Netherlands Fax 925 603 8080 Pan Cytokeratin Plus AE1 AE3 8 18 Concentrated and Prediluted Cocktail Ant
6. r GL Lipid cell steroid cell tumor of the ovary immunophenotype with analysis of potential pitfall due to endogenous biotin like activity Int J Gynecol Pathol 1995 Oct 14 4 331 8 2 Bunton TE The immunocytochemistry of cytokeratin in fish tissues Vet Pathol 1993 Sep 30 5 418 25 3 Sorensen SC et al Structural distinctions among human breast epithelial cells revealed by the monoclonal antikeratin antibodies AEI and AE3 J Pathol 1987 Oct 153 2 151 62 4 Pinkus GS Etheridge CL O Connor EM Are keratin proteins a better tumor marker than epithelial membrane antigen A comparative immunohistochemical study of various paraffin embedded neoplasms using monoclonal and polyclonal antibodies Am J Clin Pathol 1986 Mar 85 3 269 77 5 Pinkus GS et al Optimal immunoreactivity of keratin proteins in formalin fixed paraffin embedded tissue requires preliminary trypsinization An immunoperoxidase study of various tumours using polyclonal and monoclonal antibodies J Histochem Cytochem 1985 May 33 5 465 73 6 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 7 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 aoa Biocare Medical 4040 Pike Lane CE Concord CA 94520
7. the past AE1 AE3 has had problems marking certain tissues types and adenocarcinomas The addition of CK 8 18 remedies some of these problems For example a study of twenty eight lipid cell steroid cell tumors of the ovary were studied by immunohistochemistry 46 were positive for Cytokeratin 8 18 antibody 37 were positive with the Cytokeratin cocktail AE1 AE3 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 mouse and rat Clone AE1 AE3 5D3 Isotype IgG1 Total Protein Concentration 10 mg ml Call for lot specific Ig concentration Epitope Antigen AE1 AE3 CK8 18 Cellular Localization Cytoplasmic Positive Control Skin or adenocarcinoma 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 the
8. y 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 Protocol Pretreatment may be performed by heat retrieval or enzyme digestion Heat Retrieval Method Pretreatment Solution recommended Reveal Retrieve sections under pressure using Biocare s Decloaking Chamber followed by a wash in distilled water Allow solution to cool for 10 minutes then wash in distilled water Digestion Method Digest with Trypsin enzyme for 5 minutes at 37 C or for 20 minutes at RT taal Biocare Medical 4040 Pike Lane Concord CA 94520 USA Tel 800 799 9499 l CE Page of 2 www biocare net IP 162 G10 10 ml prediluted OAI 162 T60 60 tests prediluted Ready to use N A Ready to use N A Protocol Recommendations intelliP ATH and manual use Cont d Protein Block Optional Incubate for 5 10 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 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
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