CYTONEL KIT TROUBLESHOOTING GUIDE (Cytonel
Contents
1. 50 3005609 fax 358 3 3642110 VAT number F118058489 email jilab jilab fi Problem 4 Brown background on empty glass area Cause Some slide coatings may absorb staining reagents Remedy Avoid poly L lysine coated objective slides SuperFrost Plus recommended Problem 5 Brownish background staining on tissue section Cause Insufficient washing between antibody and DAB steps Remedy Wash slides for 10 sec with manual agitation Make sure that you use PBS 0 05 Tween20 Washing buffer concentrate tablet included in the kit should be dissolved in 1 liter of distilled water Problem 6 Weak staining reaction in tissue sections stored at 20C for long periods months years Cause Freeze drying Remedy For longer periods unstained frozen sections slides need to be stored airtight and preferable at 70C Problem 7 Cytoplasmic fibers of the dendritic reticulum cells stain positively in sentinel nodes Cause Expression of cytokeratin 8 in these cells These cells are occasionally stained with Cytonel Plus pancytokeratin kit The phenomenon is well documented in the histopathology literature Remedy Keep in mind during histopathologic examination Carcinoma cells have typical round or oval morphology Cells with fiber like elongations should be ignored Remedy 2 Use Cytonel 19 which reacts only with cytokeratin 19 and not with cytokeratin 8 Jilab Inc Biokatu 6 33520 TAMPERE2. Finland 358 50 3005609 jilab jilab fi
3. JILAB INC address Biokatu 6 33520 Tampere Finland tel 358 50 3005609 fax 358 3 3642110 VAT number FI18058489 email jilab jilab fi CYTONEL KIT TROUBLESHOOTING GUIDE Cytonel Plus and Cytonel 19 Problem 1 No staining reaction seen Cause Incorrect execution of the staining protocol Remedy Make sure that you follow the staining protocol steps in correct order Performance of the kit can be verified with the control slides included in the demo kit slides available also at request Compare your staining results of the control slide with the images below Problem 2 Too weak brown staining reaction Cause 1 Slide too wet when adding DAB Remedy Dry blot slide properly or as recommended use Slide Spinner Cause 2 Temperature of the thermal plate too low Remedy Adjust thermal plate to 42C Problem 3 Unwanted cells macrophages display strong cytoplasmic staining see image Cause Insufficient blocking of endogenous peroxidase activity Remedy Make sure that you prepared the fixative working solution as instructed Add 1 ml of 3 H202 and 0 5 ml blocking agent included in the kit in 100ml of commercial grade acetone Use fixative within 6 hours Please note that 30 stock solution of H202 may precipitate in the fixative Jilab Inc Biokatu 6 33520 TAMPERE Finland 358 50 3005609 jilab jilab fi JILAB INC address Biokatu 6 33520 Tampere Finland tel 358
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