Lipid Staining Kit (ST-R100) - Zen
Contents
1. zenbio p gt Lipid Staining Kit 200 Point Assay Kit in 96 well Format Cat ST R100 INSTRUCTION MANUAL 2ZBMO0012 01 STORAGE CONDITIONS e Reagents amp Buffers Room temperature 25 C For in vitro Use Only For Research Use Only Not For Use In Diagnostic Procedures LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this product No other warranties of any kind expressed or implied including without limitation implied warranties of merchantability or fitness for a particular purpose are provided by Zen Bio Inc Zen Bio Inc shall have no liability for any direct indirect consequential or incidental damages arising out of the use the results of use or the inability to use this product ORDERING INFORMATION AND TECHNICAL SERVICES Zen Bio Inc e 3200 Chapel Hill Nelson Blvd Suite 104 e PO Box 13888 e Research Triangle Park NC 27709 e Telephone 919 547 0692 e Facsimile FAX 919 547 0693 e Toll Free 1 866 ADIPOSE 866 234 7673 e Electronic mail e mail information zen bio com e World Wide Web http www zen bio com Rev 8 18 2008 Page 1 of 2 INTRODUCTION This protocol is designed to stain cells in a 96 well format using Oil Red O Oil Red O is a fat soluble diazo dye that stains neutral triglycerides and lipids For other formats please adjust the volumes added to each well according to the surface area of the well flask you are using ITEMS INCLUDED IN THE KIT Fixa2. er paper is effective it just takes longer to filter Polycarbonate filters will dissolve in isopropanol DO NOT USE THEM Remove all the Fixative Solution Wash cells at least twice with 200 ul well dH2O using a multichannel pipette Remove all the water using a multichannel pipette Make sure all the water is removed BEFORE proceeding Add 50 ul well Working Oil Red O solution WOROS at room temperature for 10 15 minutes Be careful not to touch the sides of the wells the pipette tips should go at a 90 angle into the wells Remove all of the Oil Red O solution from each well Wash 2 3 times with 60 80 ul well dHoO Pictures may be taken at this time Leave water inside wells during photography Rev 8 18 2008 Page 2 of 2
3. tive Solution Store at room temperature preferably in a cabinet approved 40 ML 1 SAS er sirage ot tammano cremea some Oil Red O Stock Solution Store at room temperature in a dark bottle preferably in a 6 ML 1 POTOSI cain approved r gorago oi Tarman cremea eomme For multi channel pipettes clear polyvinyl Other equipment reagents required but not provided with the kit Mature adipocytes or other lipid containing cells to be stained Deionized or distilled water AH920 Multi channel Pipet single channel pipette and pipette tips Tubes for preparing working solutions ASSAY PROCEDURE 96 well plate format Protocol Remove row by row all the liquid and add 100ul well Fixative Solution Incubate at room temperature for 5 minutes Repeat liquid exchange with another 100 uI well Fixative Solution Fix cells at least 1 hour at 4 C Alternatively the plate of cells in Fixative Solution may be sealed using sealing film foil not included and stored at 4 C for 7 days before proceeding Prepare Working Oil Red Solution WOROQOS Prepared fresh on day of assay 40 water 60 Oil Red O stock a Add 6 0 ml of Oil Red O Stock Solution to 4 0 ml of water and mix by inversion b This working solution must be kept at room temperature at least 20 minutes before filtering Filter out particulate material using a 1um polypropylene PTFE syringe filter Supplied with the kit or 1 Whatman filter paper not provided NOTE Filt
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