CHO HCP ELISA Kit
Uburyo bumwe bwo kwirinda immunosorbent ELISA bukoreshwa muriki kibazo.Ingero zirimo CHOK1 HCP icyarimwe zifata hamwe na HRP yanditseho ihene irwanya CHOK1 na antibody anti-CHOK1 yometse ku isahani ya ELISA, amaherezo ikora sandwich igizwe na antibody yo mu cyiciro gikomeye-HCP yanditseho antibody.Antigen-antibody idahambiriwe irashobora gukurwaho no koza isahani ya ELISA.TMB substrate yongewe kumariba kugirango reaction ihagije.Iterambere ryamabara rihagarikwa nyuma yo kongeramo igisubizo, hamwe na OD cyangwa kwinjiza agaciro k'igisubizo cya reaction kuri 450 / 650nm isomwa numusomyi wa microplate.Agaciro ka OD cyangwa agaciro kinjira kagereranijwe nibiri muri HCP mubisubizo.Kuva aho, kwibanda kwa HCP mubisubizo birashobora kubarwa ukurikije umurongo usanzwe.
Gusaba
Iki gikoresho gikoreshwa mukumenya muburyo bukubiye muri CHOK1 yakira selile isigara ya selile.
CIbigize
| S / N. | Ibigize | Kwibanda | Ububiko |
| 1 | CHOK1 HCP Igipimo | 0.5mg / mL | ≤ - 20 ℃ |
| 2 | Kurwanya CHO HCP-HRP | 0.5mg / mL | ≤ - 20 ℃, kurinda urumuri |
| 3 | TMB | NA | 2-8 ℃, urinde urumuri |
| 4 | 20 × PBST 0.05% | NA | 2-8 ℃ |
| 5 | Hagarika igisubizo | NA | RT |
| 6 | Abacuruzi ba Microplate | NA | RT |
| 7 | BSA | NA | 2-8 ℃ |
| 8 | Amashanyarazi menshi mbere yo gutwikira | NA | 2-8 ℃ |
Ibikoresho Birakenewe
| Ibikoreshwa / ibikoresho | Inganda | Cataloge |
| Umusomyi wa Microplate | Ibikoresho bya molekulari | Spectra Max M5, M5e, cyangwa bihwanye |
| Thermomixer | Eppendorf | Eppendorf / 5355, cyangwa bihwanye |
| Imvange ya Vortex | IKA | MS3 Digital, cyangwa ibisa nayo |
Ububiko n'umutekano
1.Ubwikorezi kuri -25 ~ -15 ° C.
2.Imiterere yo kubika nkuko bigaragara mu mbonerahamwe ya 1;ibice 1-2 bibitswe ≤ - 20 ° C, 5-6 bibitswe RT, 3 、4、7、8 bibitswe kuri 2-8 ℃;igihe cyemewe ni amezi 12.
Ibipimo byibicuruzwa
1.Ibyiyumvo: 1ng / mL
2.Urutonde rwo kumenya: 3- 100ng / mL
3.Icyitonderwa: Imbere-isuzuma CV≤ 10%, hagati ya CV≤ 15%
4.Ubwishingizi bwa HCP:> 80%
5.Umwihariko: Iki gikoresho ni rusange kuko gikora neza na CHOK1 HCP idashingiye kubikorwa byo kwezwa.
Witegure neza
1.PBST 0.05%
Fata ml 15 ya 20 × PBST 0,05%, ivanze muri ddH2O, kandi bigera kuri 300 ml.
2.1.0% BSA
Fata 1g ya BSA mu icupa hanyuma uyungurura muri ml 100 ya PBST 0,05%, vanga neza kugeza ushonga burundu, hanyuma ubike kuri 2-8 ° C.Buffer yateguwe ya dilution yemewe muminsi 7.Birasabwa kwitegura nkuko bikenewe.
3.Igisubizo cyo kumenya 2μg / mL
Fata 48μL ya 0.5 mg / mL Anti-CHO HCP-HRP hanyuma uyunguruze muri 11,952μL ya 1% BSA kugirango ubone igisubizo cyanyuma cya 2μg / mL igisubizo.
4.QC no Gutegura Ibipimo bya CHOK1 HCP
| Itiyo Oya. | Umwimerere | Kwibanda | Umubumbe | 1% BSA | Umubumbe wose | Final |
| A | Bisanzwe | 0.5mg / mL | 10 | 490 | 500 | 10,000 |
| B | A | 10,000 | 50 | 450 | 500 | 1.000 |
| S1 | B | 1.000 | 50 | 450 | 500 | 100 |
| S2 | S1 | 100 | 300 | 100 | 400 | 75 |
| S3 | S2 | 75 | 200 | 175 | 375 | 40 |
| S4 | S3 | 40 | 150 | 350 | 500 | 12 |
| S5 | S4 | 12 | 200 | 200 | 400 | 6 |
| S6 | S5 | 6 | 200 | 200 | 400 | 3 |
| NC | NA | NA | NA | 200 | 200 | 0 |
| QC | S1 | 100 | 50 | 200 | 250 | 20 |
Imbonerahamwe: Gutegura QC nubuziranenge
Suzuma inzira
1.Tegura reagent nkuko bigaragara muri "Reagent Preparation" hejuru.
2.Fata 50μL yubuziranenge, ingero na QCs (reba Imbonerahamwe 3) muri buri riba, hanyuma ongeramo 100μL yumuti wo kumenya (2μg / mL);Gupfundikira isahani hamwe na kashe, hanyuma ushire isahani ya ELISA kuri thermomixer.Incubate kuri 500rpm, 25 ± 3 ℃ kumasaha 2.
3.Hindura microplate muri sink hanyuma ujugunye igisubizo.Pipette 300μL ya PBST 0,05% muri buri riba kugirango woze isahani ya ELISA hanyuma ujugunye igisubizo, hanyuma usubire gukaraba inshuro 3.Hindura isahani hejuru yigitambaro gisukuye hanyuma ukame.
4.Ongeramo 100μL ya substrate ya TMB (reba Imbonerahamwe 1) kuri buri riba, funga isahani ya ELISA, hanyuma ushire mu mwijima kuri 25 ± 3 ℃ kuminota 15.
5.Pipette 100μL yo guhagarika igisubizo muri buri riba.
6.Gupima kwinjiza kumuraba wa 450 / 650nm hamwe numusomyi wa microplate.
7.Gisesengura amakuru ya SoftMax cyangwa software ihwanye.Tegura umurongo usanzwe ukoresheje uburyo bune bwo kugereranya ibintu.
Urugero rusanzwe
ICYITONDERWA: Niba kwibumbira hamwe kwa HCP murugero birenze urugero rwo hejuru rwumurongo usanzwe, bigomba kuvangwa neza na buffer ya dilution mbere yo kwipimisha.
ICYITONDERWA
Igisubizo cyo guhagarika ni 2M acide sulfurike, nyamuneka witonde kugirango wirinde kumeneka!
