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☷Y)}zק3>nr+I{M9^x}Hyw6[M::*r׶n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤlzr=Vv=ks&m.t;[M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I{Mw?r{]í8v{rM\ne7iv8.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I{Mw?r{]í8v{rM\ne7iv8.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7ioSjܼn֩~ ^}~LygsݤSkO6~T6[Mknkuemru+]í8v{>Nz{˭&5p C}z^yr+I[rS%6[M:Pm/nr+I[rӫv/|nr+I[sS)=n֩5z}:r=VvNSȥw}˭&mZ]_OG*w]˭&mZ=:3:;Se\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I{Mw?r{]í8v{rM\ne7iv8.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I{Mw?r{]í8v{rM\ne7iv8.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I{Mw?r{]í8v{rM\ne7iv8.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT?O"ccruN^w߸pR)npI[r͝}c{<$m.u*>}LVVvf=ڗr@~{6[٭8v{r91WCr+I{M9pCZm6[M:}:9x+\ne7iTnwo7w~`sݤS۫t^VvN^Pק/u*CP>L\n֩ksw{>=}j\n֩ktgۧǺOu]M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n^l6wOqcrp+yݞkw6[Mk΁˭&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݗ<.w 7iT.2os߸;P5ܤSܞSѧ^}n;ԫ]֩\nO)OjseT.X{_jseT.>׫]^l6wOqܧ'`~_)7\n1o۳7{1_*1}wV .w-[r=O^u*}skqNr{O}?{ 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xݿ]𳨥bnM͊"M$0Sȏbn$HIYU]DSE"TiBn~O539w37<ϙy=;{晙ͯ_ݛ>y7o~fi'6/~?ݟo>vG/yqCǚŶy_l_v{+/~r2"""W/~?}~OH|߼}뭷t}{w*""r|䥌|3Od韱ƌ5b'o?k\y>m^3[~pwƟ6Gwowc51qcϿcmq3c|wswes:m1s4qq<̌05o9gc>}gZvcPmxs|s8sck"/c]ƾ̵e%y죹Ό}umn7c-r\s9yss|G39;:{|7wwxcc{Ͼ}Cs{2o>;oc[r=1Gת"/˵asȾo>ݷsZ˵zd_c;sq;svr-U6_3Gs[X9ܾ{e._t}/忏ڗds1zeEgܖ[gg,͜SG۞SC5:ue}sNyri:2;^|1\o0ߟ=|nok5}Ү5ᇬ:sc~^,aε~Egާ 3t꾹=qYfvO{y1=}w+"쮿#ќ!s=5~~u>?42yZ;yksyzl^;{c>t^c;eX~6vyonww~z_w?cP(˹sDDffnjjws}Zo4|]7?{.>2 ϊX{X}y1q9yew?t}<9wevǴCߥy,g}n~evf>X?4=.s ߥٝcPs};?O0cu|s"Ԍ5T9ZޥjNXsוΣ+G|流S3z$e7׾ޫNf}3;emED.9}ε1d}3Y/}cd_i|dgXr8>=}Jnr+I{Mwhڗrߩ//o؍˭V3[q{קOq.n^}챖zO.nҾN=}TpݤVvwS^n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n^l6wOqcrp+yݞkw6[Mk΁˭&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n^l6wOqcrp+yݞkw6[Mk΁˭&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n^l6wOqcrp+yݞkw6[Mk΁˭&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iݷ߹ퟗ/VvN}~͓Tn}7isݤSeIe.}=ܤVvNEiW/y}ܤVvNOz&m.SS%nҾNtn/{)6[Mܩ?{xtv\r/r+I;u^=UxKnҾNyWO٧#:^nr+I{Mw?v k_b٥8f.w 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C}z^yr+I[rS%6[M:Pm/nr+I[rӫv/|nr+I[sS)=n֩5z}:r=VvNSȥw}˭&mZ]_OG*w]˭&mZ=:3:;Se\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I{Mw?r{]í8v{rM\ne7iv8.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I{Mw?r{]í8v{rM\ne7iv8.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I{Mw?r{]í8v{rM\ne7iv8.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT?O"ccruN^w߸pR)npI[r͝}c{<$m.u*>}LVVvf=ڗr@~{6[٭8v{r91WCr+I{M9pCZm6[M:}:9x+\ne7iTnwo7w~`sݤS۫t^VvN^Pק/u*CP>L\n֩ksw{>=}j\n֩ktgۧǺOu]M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n^l6wOqcrp+yݞkw6[Mk΁˭&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݗ<.w 7iT.2os߸;P5ܤSܞSѧ^}n;ԫ]֩\nO)OjseT.X{_jseT.>׫]^l6wOqܧ'`~_)7\n1o۳7{1_*1}wV .w-[r=O^u*}skqNr{O}?{ n֩\nOߣ&mt_~ݤSܞ4ܮn֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[Mkn7.c\n1o۳7{nr+I{M9pUܤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[Mkn7.c\n1o۳7{nr+I{M9pUܤVvNr{I=7_&:&m.o}gvFr=ݤ]ܞҫ9pUܤקOskw6{_>W+9^5cZu.u,.ݤ>OӫrM\ZN=v8.5]~u*{n7isk>SsI=4;{nr9{nr݄˽v7isvG_>9pUܤVvNr{I˭&mt6[M:&m.t7So\.ǘ5܊cngo.ݤVvnsrI˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7in6޸\n1skǼn\I˭&5Vq6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7in6޸\n1skǼn\I˭&5Vq6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.u*M\ne7iT.n֩\nO7isݤSܞnr+I[r=ݤVvNr{I˭&mt6[M:&m.o/ۿ$ PrYKSKS.{&#ll8@4t4d(M$hO,|SrrT.6q FOR RESEARCH USE ONLY Duck immunoglobulin A Drug Names Generic NameDuck immunoglobulin A (IgA)ELISA Kit. Purpose This kit allows for the determination of IgA concentrations in Duck serum, blood plasma, and other biological fluids. Principle of the assay The kit assay Duck IgA level in the sample use Purified Duck IgA antibody to coat microtiter plate wells, make solid-phase antibody, then add IgA to wells, Combined IgA 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 spectrophotometrically at a wavelength of 450 nm. The concentration of IgA in the samples is then determined by comparing the O.D. of the samples to the standard curve. Materials provided with the kit Materials provided with the kit48determinations96 determinationsStorageUser manual11Closure plate membrane22Sealed bags11Microelisa stripplate112-8!Standard720ng/ml0.5ml1 bottle0.5ml1 bottle2-8!Standard diluent1.5ml1 bottle1.5ml1 bottle2-8!HRP-Conjugate reagent3ml1 bottle6ml1 bottle2-8!Sample diluent3ml1 bottle6ml1 bottle2-8!Chromogen Solution A3ml1 bottle6ml1 bottle2-8!Chromogen Solution B3ml1 bottle6ml1 bottle2-8!Stop Solution3ml1 bottle6ml1 bottle2-8!wash solution20ml20 fold 1bottle20ml30 fold 1bottle2-8!Specimen requirements serum- coagulation at room temperature 10-20 mins centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again. plasma-use suited 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, If precipitation appeared, Centrifugal again. Urine-collect sue a sterile container, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again. The Operation of Hydrothorax and cerebrospinal fluid Reference to it. cell culture supernatant-detect secretory components, collect sue a sterile container, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant,detect the composition of cells, Dilut cell suspension with PBSPH7.2-7.4 , Cell concentration reached 1 million / ml, repeated freeze-thaw cycles, damage cells and release of intracellular components, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant, If precipitation appeared, Centrifugal again. Tissue samples- After cutting samples, check the weight,add PBSPH7.2-7.4 , Rapidly frozen with liquid nitrogen, maintain samples at 2-8! after melting,add PBSPH7.4 , Homogenized by hand or Grinders, centrifugation 20-min at the speed of 2000-3000 r.p.m. remove supernatant. extract as soon as possible after Specimen collection,and according to the relevant 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. Can t detect the sample which contain NaN3, because NaN3 inhibits HRP active. Assay procedure 1.Dilute and add sample to Standard: set 10 Standard wells on the ELISA plates coated, add Standard 100l to the first and the second well, then add Standard dilution 50l to the first and the second well, mix; take out 100l form the first and the second well then add it to the third and the forth well separately. then add Standard dilution 50l to the third and the forth well ,mix ; then take out 50l from the third and the forth well discard, add 50l to the fifth and the sixth well ,then add Standard dilution 50l to the fifth and the sixth well, mix ; take out 50l from the fifth and the sixth well and add to the seventh and the eighth well, then add Standard dilution 50l to the seventh and the eighth well ,mix ; take out 50l from the seventh and the eighth well and add to the ninth and the tenth well, add Standard dilution 50l to the ninth and the tenth well, mix , take out 50l from the ninth and the tenth well discard(add Sample 50l to each well after Diluting ,(density: 480ng/ ml 320ng/ ml 160ng/ ml 80ng/ ml 40ng/ ml) 2.add sampleSet blank wells separately (blank comparison wells don t add sample and HRP-Conjugate reagent, other each step operation is same). testing sample well. add Sample dilution 40l to testing sample well, then add testing sample 10l (sample final dilution is 5-fold), add sample to wells , don t touch the well wall 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 distilled water and reserve. 5.washingUncover Closure plate membrane, discard Liquid, dry by swing, add washing buffer to every well, still for 30s then drain, repeat 5 times, dry by pat. 6.add enzymeAdd HRP-Conjugate reagent 50l to each well, except blank well. 7.incubateOperation with 3. 8.washingOperation with 5. 9.colorAdd Chromogen Solution A 50ul and Chromogen Solution B to each well, evade the light preservation for 15 min at 37! 10.Stop the reactionAdd Stop Solution50l to each well, Stop the reaction(the blue color change to yellow color). 11.assaytake blank well as zero , Read absorbance at 450nm after Adding Stop Solution and within 15min. Important notes 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. washing buffer will Crystallization separation, it can be heated the water helps dissolve when dilute . Washing does not affect the result. add Sample with sampler Each step, And proofread its accuracy frequently, avoids the experimental error. add sample within 5 mins, if the number of sample is much , recommend to use Volley . if the testing material content is excessively higher (The sample OD is bigger than the first standard well ),please dilute Sample (n-fold), Please diluente and multiplied by the dilution factor.n5 . Closure plate membrane only limits the disposable use, to avoid cross-contamination. The substrate evade the light preservation. 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