CHO HCP ELISA Kit
Indlela ye-ELISA enenyathelo elinye i-immunosorbent isetyenziswa kolu vavanyo.Iisampulu eziqulathe i-CHOK1 HCP ngaxeshanye zisabela nge-anti-CHOK1 ebhalwe ibhokhwe echasene ne-CHOK1 kunye ne-anti-CHOK1 antibody egqunywe kwipleyiti ye-ELISA, ekugqibeleni yenze isangweji eyinkimbinkimbi yenqanaba eliqinileyo le-antibody-HCP-elibhalwe i-antibody.I-antigen-antibody engabotshwanga inokususwa ngokuhlamba ipleyiti ye-ELISA.I-substrate ye-TMB yongezwa kwiqula ukwenzela ukusabela okwaneleyo.Ukuphuhliswa kombala kumisiwe emva kokongeza isisombululo sokumisa, kunye ne-OD okanye ixabiso le-absorbeance yesisombululo sokuphendula kwi-450 / 650nm ifundwa kunye nomfundi we-microplate.Ixabiso le-OD okanye ixabiso lokuxhamla lilingana nomxholo we-HCP kwisisombululo.Ukusuka kule nto, i-HCP yoxinaniso kwisisombululo ingabalwa ngokwe-curve standard.
Isicelo
Le khithi isetyenziselwa ukufumanisa ngokobungakanani umxholo we-CHOK1 i-host host cell residues kwiisampuli.
Cabachasi
| S/N | Icandelo | Ukugxininisa | Iimeko zokuGcina |
| 1 | CHOK1 HCP Umgangatho | 0.5mg/mL | ≤–20℃ |
| 2 | I-Anti-CHO HCP-HRP | 0.5mg/mL | ≤–20℃, khusela ekukhanyeni |
| 3 | TMB | NA | 2-8℃, khusela ekukhanyeni |
| 4 | 20 × PBST 0.05% | NA | 2-8℃ |
| 5 | Misa isisombululo | NA | RT |
| 6 | Microplate sealers | NA | RT |
| 7 | BSA | NA | 2-8℃ |
| 8 | I-adsorption ephezulu yangaphambili yokugquma iiPlates | NA | 2-8℃ |
Izixhobo ezifunekayo
| Izinto ezisetyenziswayo / Izixhobo | Ukwenziwa | Ikhathalogu |
| Umfundi weMicroplate | Izixhobo zeemolekyuli | ISpectra Max M5, M5e, okanye ilingana |
| Thermomixer | Eppendorf | Eppendorf/5355, okanye efanayo |
| Umxube weVortex | IKA | MS3 Digital, okanye efanayo |
Ukugcinwa kunye nokuzinza
1.Ezothutho ku -25~-15°C.
2.Iimeko zokugcina njengoko kubonisiwe kwiThebhile 1;amacandelo 1-2 agcinwa ≤–20 °C,5-6 zigcinwa RT,3,I-4, 7,8 igcinwe kwi-2-8℃;ixesha lokuqinisekisa ziinyanga ezili-12.
Iiparamitha zemveliso
1.Uvakalelo: 1ng/mL
2.Uluhlu lokufumanisa: 3- 100ng / mL
3.Ukuchaneka: I-Intra-assay CV≤ 10%, inter- assay CV≤ 15%
4.Ukufikelela kwi-HCP: >80%
5.Ukucaciswa: Le khithi yendalo yonke njengoko iphendula ngokuthe ngqo nge-CHOK1 HCP ezimeleyo kwinkqubo yokucoca.
Ukulungiswa kwe-reagent
1.PBST 0.05%
Thatha i-15 ml ye-20×PBST 0.05%, ihlanjululwe kwi-ddH2O, kwaye yenziwe ukuya kwi-300 ml.
2.1.0% BSA
Thatha i-1g ye-BSA kwibhotile kwaye uhlambulule kwi-100 ml ye-PBST 0.05%, xuba kakuhle de uchithe ngokupheleleyo, kwaye ugcine kwi-2-8 ° C.Isithinteli sokuthambisa esilungisiweyo sisebenza kangangeentsuku ezisi-7.Kucetyiswa ukulungiselela njengoko kufuneka.
3.Isisombululo sokufumanisa 2μg/mL
Thatha i-48μL ye-0.5 mg / mL i-Anti-CHO HCP-HRP kwaye uhlambulule kwi-11,952μL ye-1% ye-BSA ukuze ufumane i-concentration yokugqibela ye-2μg / mL isisombululo sokufumanisa.
4.I-QC kunye noLungiselelo lweMigangatho ye-CHOK1 HCP
| Umbhobho Hayi. | Eyoqobo | Ukugxininisa | Umthamo | 1% BSA | Umthamo opheleleyo | Okokugqibela |
| A | Umgangatho | 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 |
Itheyibhile: Ukulungiswa kwe-QC kunye neMigangatho
Inkqubo yoVavanyo
1.Lungiselela ii-reagents njengoko kubonisiwe kwi-"Reagent Preparation" ngasentla.
2.Thatha i-50μL yemigangatho, iisampulu kunye ne-QCs (jonga kwiThebhile 3) kwiqula ngalinye, uze ungeze i-100μL yesisombululo se-Detection (2μg / mL);Gubungela ipleyiti nge-sealer, kwaye ubeke ipleyiti ye-ELISA kwi-thermomixer.Fudumeza nge-500rpm, 25±3℃ iiyure ezi-2.
3.Guqula i-microplate kwi-sink kwaye ulahle isisombululo sokugubungela.I-Pipette 300μL ye-PBST 0.05% kwiqula ngalinye ukuhlamba ipleyiti ye-ELISA kwaye ulahle isisombululo, kwaye uphinde uhlambe amaxesha ama-3.Guqula ipleyiti kwitawuli yephepha ecocekileyo kwaye wome.
4.Yongeza i-100μL ye-TMB substrate (bona iTheyibhile 1) kwiqula ngalinye, tywina ipleyiti ye-ELISA, kwaye ufukame ebumnyameni kwi-25±3℃ i-15 min.
5.I-Pipette 100μL yesisombululo sokumisa kwiqula ngalinye.
6.Ukulinganisa ukutsalwa kwe-wavelength ye-450 / 650nm kunye nomfundi we-microplate.
7.Hlalutya idatha ngeSoftMax okanye isoftware efanayo.Yila igophe elisemgangathweni ngokusebenzisa imodeli yohlengahlengiso yeparamitha ezine.
Umzekelo weCwave eMgangathweni
QAPHELA: Ukuba ukuxinwa kwe-HCP kwisampulu kudlula umda ophezulu we-curve esemgangathweni, kufuneka ihlanjululwe ngokufanelekileyo nge-dilution buffer phambi kovavanyo.
AMANQAKU
Isisombululo sokumisa yi-2M sulfuric acid, nceda uphathe ngononophelo ukuphepha ukutshiza!
