大鼠甘油三酯(TG)英文说明书

大鼠甘油三酯(TG)英文说明书 Rat triglyceride (TG) FOR RESEARCH USE ONLY --上海谷研生物公司专业代理销售 Assay range:1.5nmol/L - 40nmol/L 48 determinations Purpose This kit al ows for the determination of TG concentrations in Rat serum, cel culture supernates and other biological fluids Principle of the assay The kit assay Rat TG level in the sample, use Purified Rat TG antibody to coat microtiter plate wel s, make solid-phase antibody, then add TG to wel s, Combined TG antibody which With HRP labeled, become antibody - antigen - enzyme-antibody complex, after washing Completely, Add TMB substrate solution,TMB substrate becomes blue color At HRP enzyme-catalyzed, reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrical y at a wavelength of 450 nm. The concentration of Rat TG in the samples is then determined by comparing the O.D. of the samples to the standard curve. Materials provided with the kit 1 wash solution 20ml×1bottle 7 Stopp Solution 3ml×1 bottle Standard 2 HRP-Conjugate reagent 3ml×1 bottle 8 0.5ml×1 bottle (80nmol/L) 3 Microelisa stripplate 12wel ×4strips 9 Standard diluent 1.5ml×1bottle 4 Sample diluent 3ml×1 bottle 10 Instruction 1 Closure plate 2 5 Chromogen Solution A 3ml×1 bottle 11 membrane 1 Sealed bags 6 Chromogen Solution B 3ml×1 bottle 12 Specimen requirements 1. extract as soon as possible after Specimen col ection,and according to the relevant 1 literature, and should be experiment as soon as possible after the extraction. If it can’t, specimen can be kept in -20 ? to preserve, Avoid repeated freeze-thaw cycles. 2. Can’t detect the sample which contain NaN3, because NaN3 inhibits HRP active. Assay procedure 1. Dilute and add sample:Dilute Original density Standard as fol ow table: 40nmol/L Standard 5150µl Original density Standard+150µl Standard diluent 20nmol/L Standard 4150µl 5 Standard+150µl Standard diluent 10nmol/L Standard 150µl 4 Standard+150µl 3Standard diluent 5.0nmol/L Standard 2150µl 3 Standard +150µl Standard diluent 2.5nmol/L Standard 1150µl 2 Standard +150µl Standard diluent 2.add sample:Set blank wel s separately (blank comparison wel s don’t add sample and HRP-Conjugate reagent, other each step operation is same). testing sample wel . add Sample dilution 40µl to testing sample wel , then add testing sample 10µl (sample final dilution is 5-fold), add sample to wel s , don’t touch the wel wal as far as possible, and Gently mix. 3.Incubate: After closing plate with Closure plate membrane ,incubate for 30 min at 37?. 4.Configurate liquid: 30-fold(or 20-fold) wash solution diluted 30-fold (or 20-fold) with distil ed water and reserve. 5.washing:Uncover Closure plate membrane, discard Liquid, dry by swing, add washing buffer to every wel , stil for 30s then drain, repeat 5 times, dry by pat. 6.add enzyme:Add HRP-Conjugate reagent 50µl to each wel , except blank wel . 7.incubate:Operation with 3. 8.washing:Operation with 5. 9.color:Add Chromogen Solution A 50ul and Chromogen Solution B 50ul to each wel , evade the light preservation for 10 min at 37? 10.Stop the reaction:Add Stop Solution50µl to each wel , Stop the reaction(the blue color change to yel ow color). 11.assay:take blank wel as zero , Read absorbance at 450nm after Adding Stop Solution and within 15min. 2 Steps description Standard, Sample diluent Add Standard, Sample diluent, incubate for 30 min at 37?. Wash 5 time,Add HRP-Conjugate reagent, incubate for 30 min at 37?. Wash 5 times,Add Chromogen Solution A and B, incubate for 10 min at 37?. Add Stop Solution Read absorbance at 450nm within 15 min calculate Calculate Take the standard density as the horizontal, the OD value for the vertical ,draw the standard curve on graph paper, Find out the corresponding density according to the sample OD value by the Sample curve, multiplied by the dilution multiple, or calculate the straight line regression equation of the standard curve with the standard density and the OD value ,with the 3 sample OD value in the equation, calculate the sample density, multiplied by the dilution factor, the result is the sample actual density. Important notes 1. The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature, ELISA plates coated if has not use up after opened, the plate should be stored in Sealed bag. 2. washing buffer wil Crystal ization separation, it can be heated the water helps dissolve when dilute . Washing does not affect the result. 3. add Sample with sampler Each step, And proofread its accuracy frequently, avoids the experimental error. add sample within 5 min, if the number of sample is much , recommend to use Vol ey . 4. if the testing material content is excessively higher (The sample OD is bigger than the first standard wel ),please dilute Sample (n-fold), Please diluente and multiplied by the dilution factor.(×n×5). 5. Closure plate membrane only limits the disposable use, to avoid cross-contamination. 6. The substrate evade the light preservation. 7. Please according to use instruction strictly, The test result determination must take the microtiter plate reader as a standard. 8. Al samples, washing buffer and each kind of reject should according to infective material process. 9. Do not mix reagents with those from other lots. Storage and validity 1(Storage: 2-8?. 2(validity: six months 4