Estrogen Receptor (ER) [SP1]
Contents
1. Estrogen Receptor ER SP 1 Concentrated and Prediluted Rabbit Monoclonal Antibody Control Number 902 301 040215 Catalog Number CRM 301 A B C PRM 301 AA Description 0 1 0 5 1 0 ml concentrated 6 0 ml prediluted Dilution 1 50 1 100 Ready to use Diluent Renoir Red N A Intended Use For Research Use Only Not for use in diagnostic procedures Summary and Explanation Human estrogen receptor ER is a 66 kDa protein that acts as an estrogen dependent nuclear hormone receptor Studies have shown ER is present in the nuclei of epithelial cells in normal breast and endometrial tissues as well as a subset of breast carcinomas The SP1 clone is a high affinity rabbit monoclonal antibody directed against an epitope of the C terminus of the ER protein 1 SP1 has been shown to stain formalin fixed paraffin embedded tissues The robustness of SP1 has been demonstrated by successful immunohistochemistry using lower temperatures for antigen retrieval e g 80 or 95 C and in some instances staining can be obtained even without antigen retrieval 2 3 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 After labeling the antigen with a primary antibody an enzyme labeled polymer is then added to bind to the primary antibody This detection of the bound antibody is evidenced by a colorimetric reaction Source2. Rabbit monoclonal Species Reactivity Human others not tested Clone SP1 Isotype IgG Total Protein Concentration 10 mg ml Call for lot specific Ig concentration Epitope Antigen ER Cellular Localization Nuclear Positive Control Breast 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 Staining Protocol Recommendations manual use Peroxide Block Block for 5 minutes with Biocare s Peroxidazed 1 Pretreatment Solution recommended Diva Pretreatment Protocol Heat Retrieval Method Preheat the retrieval solution to 95 C for 30 minutes in Biocare s Decloaking Chamber Then place slides into the preheated solution and retrieve under pressure at 95 C for 40 minutes alternatively steam tissue sections for 45 60 minutes or use a water bath at 95 C for 40 minutes Allow solution to cool for 20 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 minutes at RT Probe N A Polymer Incubate fo
3. user to determine the appropriate application for its use 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 5 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 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 Technical Support Contact Biocare s Technical Support at 1 800 542 2002 for questi
4. ment M29 A4 Wayne PA 2014 Biocare Medical LLC 4040 Pike Lane Concord CA 94520 l Page 2 of 2 Tel 800 799 9499 www biocare net Fax 925 603 8080
5. ons regarding this product References 1 Cheang MC et al Immunohistochemical detection using the new rabbit monoclonal antibody SP1 of estrogen receptor in breast cancer is superior to mouse monoclonal antibody 1D5 in predicting survival J Clin Oncol 2006 Dec 24 36 5637 44 2 Rossi S et al Rabbit monoclonal antibodies a comparative study between a novel category of immunoreagents and the corresponding mouse monoclonal antibodies Am J Clin Pathol 2005 Aug 124 2 295 302 3 Cano G et al Estimation of hormone receptor status in fine needle aspirates and paraffin embedded sections from breast cancer using the novel rabbit monoclonal antibodies SP1 and SP2 Diagn Cytopathol 2003 Oct 29 4 207 11 Page 1 of 2 Fax 925 603 8080 Estrogen Receptor ER SP 1 Concentrated and Prediluted Rabbit Monoclonal Antibody Control Number 902 301 040215 References Cont d 4 Rocha R et al Rabbit monoclonal antibodies show higher sensitivity than mouse monoclonals for estrogen and progesterone receptor evaluation in breast cancer by immunohistochemstry Pathol Res Pract 2008 204 9 655 62 5 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 6 Clinical and Laboratory Standards Institute CLSI Protection of Laboratory Workers from Occupationally Acquired Infections Approved Guideline Fourth Edition CLSI docu
6. r 30 minutes at RT with a secondary conjugated 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 Biocare Medical LLC 4040 Pike Lane Concord CA 94520 coal Tel 800 799 9499 www biocare net OAR 301 T60 60 tests prediluted Ready to use N A Technical Note This antibody has been standardized with Biocare s MACH 2 detection system It can also be used on an automated staining system and with other Biocare polymer detection kits Use TBS buffer for washing steps Protocol Recommendations ONCORE Automated Slide Staining System OAR301 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 ER Rb Protocol Template Description Rb HRP Template 1 Dewaxing DS Option DS2 Antigen Retrieval AR Option AR2 low pH 95 C Reagent Name Time Temp ER Rb 30 min 25 C Limitations This product is provided for Research Use Only RUO and is not for use in diagnostic procedures Suitability for specific applications may vary and it is the responsibility of the end
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