Data Sheet
Contents
1. 1 Falini B et al ALK expression defines a distinct group of T Null lymphomas ALK lymphomas with a wide morphological spectrum Am J Pathol 1998 Sep 153 3 875 86 2 Mino Kenudson M et al A novel highly sensitive antibody allows for the routine detection of ALK rearranged lung adenocarcinomas by standard immunohistochemistry Clin Cancer Res 2010 Mar 1 16 5 1561 71 3 Paik JH et al Screening of anaplastic lymphoma kinase rearrangement by immunohistochemistry in non small cell lung cancer correlation with fluorescence in situ hybridization J Thorac Oncol 2011 Mar 6 3 466 72 4 Kim H et al Detection of ALK gene rearrangement in non small cell lung cancer a comparison of fluorescence in situ hybridization and chromogenic in situ hybridization with correlation of ALK protein expression J Thorac Oncol 2011 Aug 6 8 1359 66 5 McLeer Florin A et al Dual IHC and FISH testing for ALK gene rearrangement in lung adenocarcinomas in a routine practice a French study J Thorac Oncol 2012 Feb 7 2 348 54 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 Concor2. ALK 5A4 Concentrated and Prediluted Monoclonal Antibody ISO 9001813485 CERTIFIED Control Number 901 3041 033115 Catalog Number ACI 3041 A B API 3041 AA Description 0 1 0 5 ml concentrated 6 0 ml prediluted Dilution 1 50 1 100 Ready to use Diluent Renoir Red N A Intended Use For In Vitro Diagnostic Use ALK 5A4 is a mouse monoclonal antibody that is intended for laboratory use in the qualitative identification of aa 419 520 of ALK protein or ALK NPM chimeric 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 ALK p80 recognizes the formalin resistant epitope of native anaplastic lymphoma kinase ALK protein ALK recognizes a 200 kDa transmembrane molecule expressed only in neural tissues The ALK reacts with normal ALK protein as well as with the chimeric protein ALK NPM ALK specifically labels t 2 5 positive cells giving strong cytoplasmic staining that is also associated with nuclear staining Anaplastic large cell lymphoma ALCL is a heterogeneous group of diseases by morphology immunophenotyping and clinical presentation It can be difficult to diagnose because of its similarity to Hod
3. d 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
4. gkin s lymphoma However treatment and prognosis of Hodgkin s and ALCL is very different and it is imperative to diagnose correctly Research has also shown that a subset of lung adenocarcinomas harbor rearrangements of the ALK gene that results in the pathologic expression of a fusion protein most commonly EMLA ALK Patients with ALK rearranged lung adenocarcinomas are unresponsive to tyrosine kinase inhibitors that target EGFR but have shown dramatic improvement in response to tyrosine kinase inhibitors that target ALK in ongoing clinical trials The results from studies comparing FISH CISH and IHC were concordant The sensitivity and specificity of IHC was reported as 100 and 95 respectively Based on these findings the IHC assay using the 5A4 antibody reliably detected non small cell lung cancer with ALK rearrangement and may be useful as a screening method to identify these tumors Research has shown that ALK stains the majority of CD30 ALCL It has not been shown to stain Hodgkin s disease Reed Sternberg cells ALK should be used in a panel with CD15 CD30 TIA 1 and EMA 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 de
5. l for 10 minutes then wash in distilled water Protein Block Optional Incubate for 5 10 minutes at RT with Biocare s Background Punisher Primary Antibody Incubate for 30 60 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 Protocol Recommendations ONCORE Automated Slide Staining System OAI3041 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 ALK Protocol Template Description Ms HRP Template 1 Dewaxing DS Option DS2 Antigen Retrieval AR Option AR1 high pH 103 C Reagent Name Time Temp ALK 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 however users must validate incubation times and protocols for their specific application Use TBS buffer for washing steps Limitation
6. ns less than 0 1 sodium azide Concentrations less than 0 1 Follow the antibody specific protocol recommendations according to data sheet are not reportable hazardous materials according to U S 29 CFR 1910 1200 OSHA provided If atypical results occur contact Buiocare s Technical Support at Hazard communication and EC Directive 91 155 EC Sodium azide NaN used as a 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
7. s 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 ALK 5A4 Concentrated and Prediluted Monoclonal Antibody Control Number 901 3041 033115 Precautions Troubleshooting ISO 9001813485 CERTIFIED 1 This antibody contai
8. tection of the bound primary antibody is evidenced by an enzyme mediated colorimetric reaction Source Mouse monoclonal Species Reactivity Human others not tested Clone 5A4 Isotype IgGl Total Protein Concentration 10 mg ml Call for lot specific Ig concentration Epitope Antigen aa 419 520 of NPM ALK transcript Cellular Localization Cytoplasmic and nuclear staining dot like Positive Control Anaplastic large 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 taal Biocare Medical 4040 Pike Lane Concord CA 94520 USA Tel 800 799 9499 l CE Page of 2 www biocare net OAI 3041 T60 60 tests prediluted Ready to use N A Protocol Recommendations manual use Peroxide Block Block for 5 minutes with Biocare s Peroxidazed 1 Pretreatment Solution 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 coo
Download Pdf Manuals
Related Search