This kit is used to assay the Anti-smooth muscle antibody(ASMA)in thesample of Human’s serum, blood plasma, and other related tissue Liquid.
The kit uses a double-antibody sandwich enzyme-linked immunosorbent assay(ELISA) to assay the level of Human Anti-smooth muscle antibody(ASMA)in samples. Add Anti-smooth muscle antibody(ASMA)to monoclonal antibody Enzyme well which is pre-coated with Human Anti-smoothmuscle antibody(ASMA)monoclonalantibody, incubation; then, add Anti-smooth muscle antibody(ASMA)antibodies labeled with biotin, and combined with Streptavidin-HRP to formimmune complex; then carry out incubation and washing again to remove theuncombined enzyme. Then add Chromogen Solution A, B, the color of the liquidchanges into the blue, And at the effect of acid, the color finallybecomesyellow. The chroma of color and the concentration of the Human Substance Anti-smoothmuscle antibody(ASMA)of samplewere positively correlated.
Materials supplied in the Test Kit
1Standard(960ng/ml)0.5ml7Chromogen Solution A6ml2Standard diluent3ml8Chromogen Solution B6ml3Microelisa Stripplate12w×8s9Stop Solution6ml4Str- HRP-Conjugate Reagent6ml10Instruction1530×wash solution20ml11Closure plate membrane26Biotin-ASMA Ab1ml12Sealed bags1
Materials required but not supplied
1. 37 ℃incubator 2.Standard Enzyme reader3. Precisionpipettes and Disposable pipette tips 4. Distilled water5. Disposabletubes for sample dilution 6. Absorbent paper
1. Beeningtaken out from the 2-8℃ environment, the kit should be balanced 30 minutes inthe ambient temperature then use. If the Coated plates of Enzyme haven’t beenused up after opened, the remaining plates should be stored in Sealed bag.2. For eachstep, add Sample with sample injector which should be calibrated frequently, inorder to avoid unnecessary experimental tolerance.3. heoperation shall be carried out accordance to the instructions strictly. Andtest results must be based on the readings of the Enzyme reader.4. In order to avoid cross-contamination, it isforBIDden to re-use the suction head and seal plate membrane in your hands.5. Allsamples, washing buffer and each kind of reject should according to infectivematerial process.6. The idleagents shall be put up or covered. Do not use reagent with different batches.And use them before expired date.7. Thesubstrate B is light-sensitive. Prolonged exposure to light is forBIDden.
Manually washing method:shake awaythe remain liquid in the enzyme plates; place some bibulous papers on thetest-bed, and flap the plates on the upside down strongly. Inject at least0.35ml after-dilution washing solution into the well, and marinate 1~2 minutes.Repeat this process according to your requirements.Automatic washing method:if there isautomatic washing machine, it should only be used in the test when you arequite familiar with its function and performance.
Intra-assay Precision(Precision within an assay): 3 samples with low, middle and high level HumanASMAwere tested 20 times on one plate,respectively. Inter-assay Precision(Precision between assays): 3 samples with low, middle and high level HumanASMA were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100Intra-Assay:CV<10% Inter-Assay: CV<12%
1. Can’tdetect the sample which contain NaN3, because NaN3 inhibits HRP active.2. extract assoon as possible after Specimen collection,and according to the relevantliterature, and should be experiment as soon as possible after the extraction.If it can’t, specimen can be kept in -20 ℃ to preserve, Avoid repeatedfreeze-thaw cycles.3. serum-coagulation at room temperature 10-20 mins,centrifugation 20-min atthe speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared,Centrifugal again.4.plasma-usesuited EDTA or citrate plasma as an anticoagulant,mix 10-20 mins,centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, Ifprecipitation appeared, Centrifugal again.5.Urine-collect sue a sterile container, centrifugation 20-min at the speed of2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugalagain. The Operation ofHydrothorax and cerebrospinal fluid Reference to it.6.cell culturesupernatant-detect secretory components, collect sue a sterile container,centrifugation 20-min at the speed of 2000-3000 r.p.m. removesupernatant,detect the composition of cells, Dilut cell suspension with PBS(PH7.2-7.4), Cellconcentration reached 1 million / ml, repeated freeze-thaw cycles, damage cellsand release of intracellular components, centrifugation 20-min at the speed of2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugalagain.7.Tissuesamples- After cutting samples, check the weight,add PBS(PH7.2-7.4), Rapidlyfrozen with liquid nitrogen, maintain samples at 2-8℃ after melting,add PBS(PH7.4), Homogenizedby hand or Grinders, centrifugation 20-min at the speed of 2000-3000 r.p.m.remove supernatant.