CHO HCP ELISA Kit
Ib kauj ruam immunosorbent ELISA yog siv rau hauv qhov kev xeem no.Cov qauv uas muaj CHOK1 HCP ib txhij hnov mob nrog HRP-labeled tshis anti-CHOK1 antibody thiab anti-CHOK1 antibody coated rau ntawm ELISA phaj, thaum kawg tsim ib qho nyuaj-theem antibody-HCP-labeled antibody.Unbound antigen-antibody tuaj yeem raug tshem tawm los ntawm kev ntxuav lub phaj ELISA.TMB substrate yog ntxiv rau lub qhov dej kom txaus cov tshuaj tiv thaiv.Kev txhim kho xim yog nres tom qab ntxiv cov tshuaj nres, thiab OD lossis absorbance tus nqi ntawm cov tshuaj tiv thaiv ntawm 450/650nm yog nyeem nrog tus nyeem ntawv microplate.Tus nqi OD lossis tus nqi absorbance yog proportional rau HCP cov ntsiab lus hauv cov tshuaj.Los ntawm qhov no, HCP concentration hauv cov tshuaj tuaj yeem suav tau raws li tus qauv nkhaus.
Daim ntawv thov
Cov khoom siv no yog siv los txheeb xyuas cov ntsiab lus ntawm CHOK1 tus tswv tsev cell protein residues hauv cov qauv.
Ccov omponents
| S/N | Cheebtsam | Nco ntsoov | Kev cia khoom |
| 1 | CHOK1 HCP Txuj | 0.5 mg / ml | ≤-20 ℃ |
| 2 | Anti-CHO HCP-HRP | 0.5 mg / ml | ≤-20 ℃, tiv thaiv lub teeb |
| 3 | TMB | NA | 2-8 ℃, tiv thaiv lub teeb |
| 4 | 20 × PBST 0.05% | NA | 2-8 ℃ |
| 5 | Nres kev daws teeb meem | NA | RT |
| 6 | Microplate sealers | NA | RT |
| 7 | BSA | NA | 2-8 ℃ |
| 8 | High adsorption pre-coating phaj | NA | 2-8 ℃ |
Khoom siv yuav tsum tau
| Consumables / khoom siv | Kev tsim khoom | Catalog |
| Microplate nyeem ntawv | Molecular Devices | Spectra Max M5, M5e, los yog sib npaug |
| Thermomixer | Eppendorf | Eppendorf/5355, los yog sib npaug |
| Vortex tov khoom | IKA | MS3 Digital, los yog sib npaug |
Cia thiab ruaj khov
1.Kev thauj mus los ntawm -25 ~ -15 ° C.
2.Cia tej yam kev mob raws li qhia nyob rau hauv Table 1;Cheebtsam 1-2 yog khaws cia ≤-20 ° C, 5-6 khaws cia RT, 3,4,7,8 yog khaws cia ntawm 2-8 ℃;lub sijhawm siv tau yog 12 lub hlis.
Khoom parameters
1.rhiab heev: 1ng / ml
2.Kev kuaj pom ntau: 3-100ng / mL
3.Precision: Intra-assay CV≤ 10%, inter- assay CV≤ 15%
4.HCP kev pab:> 80%
5.Tshwj xeeb: Cov khoom siv no yog universal raws li nws tshwj xeeb reacts nrog CHOK1 HCP ywj siab ntawm purification txheej txheem.
Kev npaj reagent
1.PBST 0.05%
Noj 15 ml ntawm 20 × PBST 0.05%, diluted hauv ddH2O, thiab ua kom txog li 300 ml.
2.1.0% BSA
Siv 1g BSA los ntawm lub raj mis thiab dilute hauv 100 ml ntawm PBST 0.05%, sib tov kom txog thaum yaj tag, thiab khaws cia ntawm 2-8 ° C.Qhov kev npaj dilution tsis siv tau rau 7 hnub.Nws raug nquahu kom npaj raws li qhov xav tau.
3.Kev kuaj pom kev daws teeb meem 2μg / mL
Noj 48μL ntawm 0.5 mg / mL Anti-CHO HCP-HRP thiab dilute hauv 11,952μL ntawm 1% BSA kom tau txais qhov kawg ntawm 2μg / mL tshuaj xyuas.
4.QC thiab Kev Npaj ntawm CHOK1 HCP Cov Qauv
| Tube Tsis muaj. | Keeb kwm | Nco ntsoov | Ntim | 1% BSA | tag nrho ntim | Kawg |
| A | Txuj | 0.5 mg / ml | 10 | 490 ib | 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 |
Table: Kev Npaj QC thiab Cov Qauv
Cov txheej txheem xeem
1.Npaj cov reagents raws li qhia hauv "Reagent Preparation" saum toj no.
2.Siv 50μL ntawm cov qauv, cov qauv thiab QCs (saib rau Table 3) rau hauv txhua lub qhov dej, tom qab ntawd ntxiv 100μL ntawm Kev Ntsuas Tshuaj (2μg / mL);Npog phaj nrog sealer, thiab muab ELISA phaj rau ntawm lub thermomixer.Incubate ntawm 500rpm, 25 ± 3 ℃ rau 2 teev.
3.Hloov lub microplate hauv lub dab dej thiab pov tseg cov txheej txheem.Pipette 300μL ntawm PBST 0.05% rau hauv txhua qhov dej los ntxuav ELISA phaj thiab muab pov tseg cov tshuaj, thiab rov ua dua 3 zaug.Hloov lub phaj rau ntawm daim ntawv so tes thiab so kom qhuav.
4.Ntxiv 100μL ntawm TMB substrate (saib Daim duab 1) rau txhua qhov dej, kaw lub phaj ELISA, thiab ua rau hauv qhov tsaus ntuj ntawm 25 ± 3 ℃ rau 15 min.
5.Pipette 100μL ntawm cov tshuaj nres rau hauv txhua qhov dej.
6.Ntsuas absorbance ntawm wavelength ntawm 450/650nm nrog microplate nyeem ntawv.
7.Txheeb xyuas cov ntaub ntawv los ntawm SoftMax lossis sib npaug software.Daim phiaj txheem nkhaus los ntawm kev siv plaub-parameter logistic regression qauv.
Standard Curve Piv txwv
CEEB TOOM: Yog hais tias qhov concentration ntawm HCP nyob rau hauv tus qauv tshaj qhov siab tshaj qhov txwv ntawm tus qauv nkhaus, nws yuav tsum tau kom zoo diluted nrog dilution buffer ua ntej kuaj.
CEEB TOOM
Cov tshuaj nres yog 2M sulfuric acid, thov ua tib zoo saib xyuas kom tsis txhob txaws!
