CHO HCP ELISA Kit
O le tasi-step immunosorbent ELISA metotia o loʻo faʻaaogaina i lenei suʻega.O fa'ata'ita'iga o lo'o iai CHOK1 HCP fa'atasi fa'atasi ma le 'oti fa'aigoa HRP anti-CHOK1 antibody ma le anti-CHOK1 antibody fa'apipi'iina i luga o le ipu ELISA, mulimuli ane fa'atupuina se sanuisi lavelave o le antibody fa'aigoa-HCP fa'ailoga.E mafai ona aveese le antigen-antibody e le fusifusia e ala i le fufuluina o le ipu ELISA.E fa'aopoopo le mea'ai TMB i le vaieli ina ia lava le tali.O le atinaʻeina o lanu e taofi pe a uma ona faʻaopoopo le fofo taofi, ma o le OD poʻo le faʻaogaina o le tau o le tali tali i le 450 / 650nm e faitau ma se microplate faitau.O le tau o le OD poʻo le faʻafefeteina e faʻatusatusa i le HCP i totonu o le fofo.Mai lenei mea, o le faʻaogaina o le HCP i le fofo e mafai ona fuafua e tusa ai ma le faʻasologa masani.
Fa'aaogāga
O lenei pusa o loʻo faʻaaogaina e suʻesuʻeina faʻatusatusa mea o loʻo i totonu ole CHOK1 host cell protein toega i faʻataʻitaʻiga.
Ctagata tetee
| S/N | Vaega | Fa'atotonuga | Tulaga Teuteu |
| 1 | CHOK1 HCP Tulaga | 0.5mg/mL | ≤–20 ℃ |
| 2 | Anti-CHO HCP-HRP | 0.5mg/mL | ≤–20℃, puipuia mai le malamalama |
| 3 | TMB | NA | 2-8 ℃, puipuia mai le malamalama |
| 4 | 20 × PBST 0.05% | NA | 2-8 ℃ |
| 5 | Taofi fofo | NA | RT |
| 6 | Fa'amau fa'amau | NA | RT |
| 7 | BSA | NA | 2-8 ℃ |
| 8 | Adsorption muamua faʻapipiʻi Papatusi | NA | 2-8 ℃ |
Meafaigaluega Manaomia
| Mea'ai/Meafaigaluega | Gaosimea | Fa'amaumauga |
| Faitautusi microplate | Meafaigaluega mole | Spectra Max M5, M5e, pe tutusa |
| Mea fa'a vevela | Eppendorf | Eppendorf/5355, pe tutusa |
| Vortex fa'afefiloi | IKA | MS3 Digital, pe tutusa |
Teuina ma le mautu
1.Felauaiga ile -25~-15°C.
2.Tulaga e teu ai e pei ona fa'aalia i le Laulau 1;vaega 1-2 o loʻo teuina ≤–20°C, 5-6 o loʻo teuina RT,3,4、7、8 o loʻo teuina ile 2-8℃;ole vaitaimi fa'ataga ole 12 masina.
Fa'atauga o oloa
1.Ma'ale'ale: 1ng/mL
2.Su'esu'ega va'aiga:3- 100ng/mL
3.Sa'o: CV≤ 10%, su'esu'e fa'atasi CV≤ 15%
4.HCP fa'asalalau: >80%
5.Fa'apitoa: O lenei pusa e fa'apitoa ona e tali fa'atasi ma CHOK1 HCP e tutoatasi mai le fa'amamāina.
Sauniuniga reagent
1.PBST 0.05%
Ave le 15 ml o le 20 × PBST 0.05%, faʻafefe i le ddH2O, ma faia e oo atu i le 300 ml.
2.1.0% BSA
Ave le 1g o le BSA mai le fagu ma faʻafefe i le 100 ml o le PBST 0.05%, faʻafefiloi lelei seia maeʻa atoa, ma teu i le 2-8°C.O le faʻamaʻi faʻafefeteina saunia e aoga mo le 7 aso.E fautuaina e sauni pe a manaʻomia.
3.Su'e fofo 2μg/mL
Ave le 48μL o le 0.5 mg / mL Anti-CHO HCP-HRP ma faʻafefe i le 11,952μL o le 1% BSA e maua ai se faʻasalaga mulimuli o le 2μg / mL faʻamaonia fofo.
4.QC ma Sauniuniga o CHOK1 HCP Tulaga
| Faila Leai. | Uluai | Fa'atotonuga | Volume | 1%BSA | Volume atoa | Mulimuli |
| A | Tulaga | 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 |
Laulau: Sauniuniga o QC ma Tulaga Fa'atonu
Fa'asologa o Su'esu'ega
1.Sauni mea fa'afo'i e pei ona fa'ailoa mai i le “Sauniuniga Fa'afofoga” i luga.
2.Ave le 50μL o faʻataʻitaʻiga, faʻataʻitaʻiga ma QCs (silasila i le Laulau 3) i totonu o vaieli taʻitasi, ona faʻaopopo lea o le 100μL o le Suʻega fofo (2μg / mL);Ufiufi le ipu i le fa'amaufa'ailoga, ma tu'u le ipu ELISA i luga o le mea fa'afefiloi.Taofi ile 500rpm, 25±3℃ mo le 2 itula.
3.Su'e le microplate i totonu o le pusa ma lafoa'i le vaifofo fa'apipi'i.Pipette 300μL o le PBST 0.05% i totonu o vaieli taitasi e fufulu ai le ipu ELISA ma lafoai le vaifofo, ma toe fai le fufuluina 3 taimi.Su'e le ipu i luga o se solo pepa mama ma fa'amago.
4.Fa'aopoopo le 100μL o le TMB substrate (silasila i le Laulau 1) i vaieli ta'itasi, fa'amaufa'ailoga le ipu ELISA, ma fa'anofo i le pogisa i le 25±3 ℃ mo le 15 min.
5.Paipa 100μL ole vaifofo taofi ile vaieli taitasi.
6.Fua le mitiia ile umi ole galu ole 450/650nm ile microplate reader.
7.Su'e fa'amaumauga e SoftMax po'o se polokalama fa'atusa.Fua le pi'o masani e ala i le fa'aogaina o se fa'ata'ita'iga fa'ata'ita'iga fa'asologa fa'asologa.
Fa'ata'ita'iga Pipi Tulaga
FAAMANATU: Afai o le maualuga o le HCP i totonu o le faʻataʻitaʻiga e sili atu i le tapulaʻa pito i luga o le taʻavale masani, e manaʻomia ona faʻamaʻa lelei i le faʻafefete faʻafefe aʻo leʻi faia suʻega.
FAAMATALAGA
Ole vaifofo taofi ole 2M sulfuric acid, faʻamolemole tago ma le faʻaeteete e aloese mai le faʻafefe!
