User Manual
Contents
1. Buffer 10ML Universal One step Indirect Enhancer 100ML ELISA Kit M 100X ELISA Probe M1 1ML With pNPP Substrate 10X Rapid Wash Solution 100ML Suitable for mouse primary oNPP Solution 100ML antibodies User Manual One copy 1 kit 1000 assays E G71 Universal One step Indirect ELISA Kit G With pNPP Substrate Suitable for goat primary antibodies 1 kit 1000 assays 10XCoating Buffer 10ML Enhancer 100ML 100X ELISA Probe G1 1ML 10X Rapid Wash Solution 100ML pNPP Solution 100ML User Manual One copy Product Description Universal One step Indirect ELISA is the latest breakthrough technology patent pending for identifying and quantifying either antigen or antibody Unlike regular classical ELISA the One step Indirect ELISA not only needs no Capture Tel 201 882 2593 Fax 201 882 2593 One step Indirect ELISA D ARE rt RAH antibody and secondary antibody but also replaces blocking primary antibody binding washing and secondary antibody binding with a rapid one step reaction Therefore using Universal M One step Indirect ELISA kit you can rapidly get your ELISA results around one hour The One step Indirect ELISA kit is a universal kit which is suitable for most primary antibodies without labeling as shown on the table above Each kit is sufficient to perform 1000 assays in 96 well plates Application e Detection and quantification of antigen e Identification2. and titration of primary antibody e Quantification of antigen specific antibody Features Easy One step reaction instead of blocking primary antibody binding washing and secondary antibody binding Rapid Get results around one hour Universal Suitable for most primary antibodies and no need of secondary antibody Reproducible Highly reproducible results Storage e Enhancer for continuous use store in 2 8 C for up to 3 months or store in 20 C for up to a year e 50X ELISA Probe for continuous use store in 20 C for up to 6 months or store in 70 C manual defrost freezer for up to a year Repeated freezing and thawing is not recommended e Rapid Wash Solution store in room temperature for up to one year Note Kit will be shipped in ambient temperature One step Indirect ELISA v s Regular Indirect ELISA ELISA Forgetit jal Antibody Titration by Two Methods o One step a Regular 0 01 Log Antibody ng ul 1505 Regents Ct Hillsborough NJ 08844 1 Tel 201 882 2593 Protocol Materials required but not included in the kit e Antigen Purified or partially purified antigen purity or gt 30 is recommended e Primary Antibody Antigen specific IgG Antibody Note We currently provide six types of Universal One step Indirect ELISA kits Kits labeled with R are suitable for rabbit primary antibodies with M suitable for mouse primary antibodies and
3. 200ul well of 1X Rapid Wash Solution Note Each plate must have certain wells coated with buffer alone as a negative control Procedure 1 One step Reaction If test samples are antibodies Dilute the sample antibody or standard antibody with Enhancer to make one or a series of dilutions make 100ul each concentration at the range of 0 1 1000ng ml for a duplicate tests Note make at least 100ul Enhancer alone as a negative control for each plate Apply 50ul each of these diluted solution or the negative control to each well of the antigen coated ELISA plate add 50ul of the above pre diluted 2X ELISA probe to each well and then incubate the plate for 40 60 minute at room temperature or overnight at 4 C with gentle shaking If test samples are antigens Mix 3 10ug of primary antibody and 200ul of the 50X ELISA Probe with 10ml Enhancer this Fax 201 882 2593 amount is sufficient for one 96 well plate Apply 100ul of this mixture to each well of the antigen coated ELISA plate and then incubate the plate for 40 60 minute at room temperature or overnight at 4 C with gentle shaking 2 Wash Wash all wells 3 times with 200ul of 1X Rapid Wash Solution it is made by diluting 100ml of the kit provided 10X Rapid Wash Solution with 900ml distilled water 3 Development e Using TMB substrate for kits IE R70 IE M70 and IE G70 Mix 100ul of 100X TMB solution A and 100ul of 100XT buffer B with 10ml of distilled water then ap
4. 4 Columbia Bio LLC WWW columbIa D10 com E mail siles icolunbia bia com User Manual Universal One step Indirect ELISA Kit For R amp D Use only Product Name amp Catalog Number Catalog Product Name Size E R70 Universal One step Indirect ELISA kKit R With TMB Substrate Suitable for rabbit primary antibodies 1 kit 1000 assays Content 10XCoating Buffer 10ML Enhancer 100ML 100X ELISA Probe RO 1ML 10X Rapid Wash Solution 100ML 100XTMB Solution Set 2ML User Manual One copy E M70 Universal One step Indirect ELISA Kit M With TMB Substrate Suitable for mouse primary antibodies 1 kit 1000 assays 10XCoating Buffer 10ML Enhancer 100ML 100X ELISA Probe MO 1ML 10X Rapid Wash Solution 100ML 100XTMB Solution Set 2ML User Manual One copy E G70 Universal One step Indirect ELISA Kit G With TMB Substrate Suitable for goat primary antibodies 1 kit 1000 assays 10XCoating Buffer 10ML Enhancer 100ML 100X ELISA Probe GO 1ML 10X Rapid Wash Solution 100ML 100XTMB Solution Set 2ML User Manual One copy E R71 Universal One step Indirect ELISA Kit R With pNPP Substrate Suitable for rabbit primary antibodies 1 kit 1000 assays 10XCoating Buffer 10ML Enhancer 100ML 100X ELISA Probe R1 1ML 10X Rapid Wash Solution 100ML pNPP Solution 100ML User Manual One copy w EMI 10XCoating
5. ply 100ul of this 1XTMB mixture to each well and wait till desired color appears Measure absorbance at 370nm without stop or at 450nm after stop the development by adding 100ul well 1N HCL or H2SO4 with a proper microplate reader e Using pNPP substrate for kits IE R71 IE M71 and IE G71 To each well add 100ul pNPP solution and wait till desired colors appear read the absorbance at 405nm with a proper microplate reader optionally stop developing by adding 100ul well 3N HCI and read the absorbance at 405nm Related Products WB R51 WB M51 WB G51 WB R50 WB M50 WB G50 Universal One step Western Blot Kits IHC R60 IHC M60 IHC G60 IHC R61 IHC M61 IHC G61 Universal One step IHC Kits Universal One step Sandwich ELISA Kits E R 3 E G73 E R74 E G74 E R80 E M80 E G80 E R81 E M81 E G81 Universal One step Cell based ELISA Kits Universal One step IgG Quantification ELISA Kits E R90 E M90 E H90 E G90 D ae 7A 1505 Regents Ct Hillsborough NJ 08844 7
6. with G suitable for goat primary antibodies e Standard if needed lf test samples are antibodies use primary antibody with a known concentration of the antigen specific IgG lf test samples are antigens use purified antigen with known concentration e Stop solution 1N HCI or H2SO4 for TMB substrate or 3N HCl or H2SO4 for pNPP substrate e Solid Phase 96 well microtiter ELISA plate Reagent Preparation If test samples are antibodies Coating plate Dissolve 50 100ug antigen with 10ml coating buffer provided in the kit to a concentration around 5 10ug ml Apply 100ul of this antigen solution to each well and incubate for two hours at room temperature or overnight at 4 C then remove the coating solution and wash wells twice with 200ul well of 1X Rapid Wash Solution 2X ELISA Probe Dilute 200ul of the 50X ELISA Probe red solution provided in the kit with 5ml Enhancer blue solution provided in the kit and mix well This amount is sufficient for one 96 well plate lf test samples are antigens Coating plate Dissolve the sample antigen to be tested or standard antigen with coating buffer provided in the kit to make one or a series of dilutions make 200ul each concentration at the range of 5 5000ng ml for a duplicate tests Apply 100ul of the diluted antigen solution to each well After incubation for two hours at room temperature or overnight at 4 C remove the coating solution and wash wells twice with
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