TTF-1 [SPT24] - Zytomed Systems
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1. G7G3 1 in LADC 72 4 and 65 4 respectively However the study demonstrated that SPT24 stained a higher percentage of lung SqCC 16 8 vs 1 Higher sensitivity of SPT24 in lung SqCC has also been shown to be heavily influenced by different detection systems 4 5 Higher sensitivity for LADC versus lung SqCC can be achieved with SPT24 compared to 8G7G3 1 while retaining specificity by the use of a cut off value and optimal antibody titer In an in house study SPT24 was titered to achieve negative staining in normal liver no cytoplasmic staining observed A cut off value of 210 of tumor cells positive for TTF 1 with a staining intensity of 2 1 was used to identify TTF 1 positive cases Using this approach SPT24 was highly sensitive for LADC 53 60 88 compared to 8G7G3 1 38 60 63 with equivalent specificity for both clones versus lung SqCC 2 137 1 5 Use of lung SqCC specific markers such as Desmoglein 3 and p40 may identify TTF 1 positive cases of squamous cell origin Additionally the use of Napsin A may confirm lung adenocarcinoma as the co expression of Napsin A and TTF 1 in lung cancers has been shown to be more pulmonary specific than either one used alone 7 Finally unlike clone 8G7G3 1 no cytoplasmic staining in lung cancers has been observed with clone SPT24 8 Principle of Procedure Antigen detection in tissues and cells is a multi step immunohistochemical process The initial step binds the primary ant2. TTF 1 SPT24 Concentrated and Prediluted Monoclonal Antibody ISO 9001813485 CERTIFIED Control Number 901 3126 011515 Catalog Number ACI 3126 A C API 3126 AA Description 0 1 1 0 ml concentrated 6 0 ml prediluted Dilution 1 100 1 200 Ready to use Diluent Renoir Red N A Intended Use For In Vitro Diagnostic Use TTF 1 SPT24 is a mouse monoclonal antibody that is intended for laboratory use in the qualitative identification of TTF 1 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 Thyroid transcription factor 1 TTF 1 is a 38 kDa member of the NKX2 family of homeodomain transcription factors TTF 1 is mostly detected in primary lung adenocarcinomas and small cell carcinomas 1 TTF 1 can be very useful in lung cancers when used in a panel with Desmoglein 3 p40 and Napsin A antibodies 2 3 Commercially available thyroid transcription factor 1 TTF 1 monoclonal antibodies 8G7G3 1 and SPT24 have been shown to have different sensitivities in lung adenocarcinomas LADC and lung squamous cell carcinomas SqCC 4 6 A study by Masai et al demonstrated that SPT24 was much more sensitive than 8
3. ibody 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 SPT24 Isotype IgGl kappa Total Protein Concentration 10 mg ml Call for lot specific Ig concentration Epitope Antigen TTF 1 Thyroid transcription factor 1 al Biocare Medical 4040 Pike Lane Concord CA 94520 usa Page of 2 Tel 800 799 9499 www biocare net OAI 3126 T70 70 tests prediluted Ready to use N A Cellular Localization Nuclear Positive Tissue Control Lung adenocarcinoma Negative Tissue Control Normal liver lung squamous cell carcinoma 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 Pret
4. ion factor 1 expression in carcinomas of the lung breast pancreas colon kidney thyroid and malignant mesothelioma Hum Pathol 2010 Jan 41 1 20 5 8 Bejarano PA Mousavi F Incidence and significance of cytoplasmic thyroid transcription factor 1 immunoreactivity Arch Pathol Lab Med 2003 Feb 127 2 193 5 9 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 10 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 EMERGO EUROPE Molenstraat 15 2513 BH The Hague The Netherlands Fax 925 603 8080
5. reatment Perform heat retrieval using Biocare s Diva Decloaker Refer to the Diva Decloaker product data sheet for specific instructions 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 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 QNCORE Automated Slide Staining System OAI3126 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 TTF 1 SPT24 Protocol Template Description Ms HRP Template 1 Dewaxing DS Option DS2 Antigen Retrieval AR Option AR1 high pH 103 C Reagent Name Time Temp TTF 1 SPT24 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 m
6. rotocol recommendations according to data sheet provided If atypical results occur contact Biocare s Technical Support at 1 800 542 2002 References 1 Di Loreto C et al Immunocytochemical expression of tissue specific transcription factor 1 in lung carcinoma J Clin Pathol 1997 Jan 50 1 30 2 2 Tacha D et al A 6 antibody panel for the classification of lung adenocarcinoma versus squamous cell carcinoma Appl Immunohistochem Mol Morphol 2012 May 20 3 201 7 3 Brown AF et al Tissue preserving antibody cocktails to differentiate primary squamous cell carcinoma adenocarcinoma and small cell carcinoma of lung Arch Pathol Lab Med 2013 Sep 137 9 1274 81 4 Masai K et al Expression of squamous cell carcinoma markers and adenocarcinoma markers in primary pulmonary neuroendocrine carcinomas App Immunohistochem Mol Morphol 2013 Jul 21 4 292 7 5 Matoso A et al Comparison of thyroid transcription factor 1 expression by 2 monoclonal antibodies in pulmonary and nonpulmonary primary tumors Appl Immunohistochem Mol Morphol 2010 Mar 18 2 142 9 6 Ord ez NG Value of thyroid transcription factor 1 immunostaining in tumor diagnosis a review and update Appl Immunohistochem Mol Morphol 2012 Oct 20 5 429 44 tal Biocare Medical 4040 Pike Lane Concord CA 94520 Tel 800 799 9499 CE USA Page 2 of 2 www biocare net References Cont d 7 Bishop JA Sharma R Illei PB Napsin A and thyroid transcript
7. ust 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 EMERGO EUROPE CE Molenstraat 15 2513 BH The Hague The Netherlands Fax 925 603 8080 TTF 1 SPT24 Concentrated and Prediluted Monoclonal Antibody ISO 9001813485 CERTIFIED Control Number 901 3126 011515 Limitations Cont d 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 Assa
8. ys 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 volumes of water to prevent azide build up in plumbing Center for Disease Control 1976 National Institute of Occupational Safety and Health 1976 9 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 10 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 Troubleshooting Follow the antibody specific p
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