CHO HCP ELISA Kit
Mokhoa o le mong oa immunosorbent ELISA o sebelisoa tekong ena.disampole tse nang le CHOK1 HCP ka nako e le nngwe di sebetsana le selwantsha-mogare sa podi ya anti-CHOK1 e ngotsweng ke HRP le selwantsha-mogare sa anti-CHOK1 se tlotsitsweng ka poleiti ya ELISA, mme qetellong ya etsa sandwich complex ya solid-phase antibody-HCP-lebelled antibody.Antigen-antibody e sa tlamisoang e ka tlosoa ka ho hlatsoa poleiti ea ELISA.Karolo e nyenyane ea TMB e kenngoa selibeng bakeng sa karabelo e lekaneng.Tsoelo-pele ea 'mala e emisoa ka mor'a ho eketsa tharollo ea ho emisa,' me boleng ba OD kapa monyetla oa tharollo ea karabelo ho 450/650nm bo baloa ka sebali sa microplate.Boleng ba OD kapa boleng ba ho monya bo lekana le litaba tsa HCP tharollong.Ho tloha ho sena, mahloriso a HCP ka tharollo a ka baloa ho ea ka sekhahla se tloaelehileng.
Kopo
Setsi sena se sebelisetsoa ho bona ka bongata boteng ba masala a protheine ea lisele tsa CHOK1 ka lisampole.
Cbahanyetsi
| S/N | Karolo | Ho tsepamisa maikutlo | Maemo a polokelo |
| 1 | CHOK1 HCP Standard | 0.5mg/mL | ≤–20℃ |
| 2 | Anti-CHO HCP-HRP | 0.5mg/mL | ≤–20℃, sireletsa khanyeng |
| 3 | TMB | NA | 2-8 ℃, sireletsa khanyeng |
| 4 | 20 × PBST 0.05% | NA | 2-8℃ |
| 5 | Emisa tharollo | NA | RT |
| 6 | Li-sealer tsa microplate | NA | RT |
| 7 | BSA | NA | 2-8℃ |
| 8 | Phahameng adsorption pele ho roala Lipoleiti | NA | 2-8℃ |
Thepa e Hlokehang
| Disebediswa/ Thepa | Tlhahiso | Catalog |
| Sebali sa microplate | Lisebelisoa tsa limolek'hule | Spectra Max M5, M5e, kapa e lekanang le eona |
| Thermomixer | Eppendorf | Eppendorf/5355, kapa e lekanang |
| Motsoako oa Vortex | IKA | MS3 Digital, kapa e lekanang |
Ho boloka le botsitso
1.Lipalangoang ho -25~-15°C.
2.Maemo a polokelo a bonts'itsoe ho Lethathamo la 1;likarolo tsa 1-2 li bolokiloe ≤-20 ° C, 5-6 li bolokiloe RT, 3,4, 7, 8 li bolokiloe ho 2-8 ℃;nako ea ho sebetsa ke likhoeli tse 12.
Lihlahisoa tsa lihlahisoa
1.Boikutlo: 1ng/mL
2.Sebaka sa ho lemoha: 3-100ng / mL
3.Ho nepahala: Intra-assay CV≤ 10%, inter-assay CV≤ 15%
4.Kakaretso ea HCP: 80%
5.Tsebo e ikhethang: Setsi sena se sebetsa hohle kaha se sebetsana ka kotloloho le CHOK1 HCP ntle le ts'ebetso ea tlhoekiso.
Boitokisetso ba reagent
1.PBST 0.05%
Nka 15 ml ea 20×PBST 0.05%, e hlapolotsoeng ka ddH2O, mme e entsoe ka 300 ml.
2.1.0% BSA
Nka 1g ea BSA ka botlolong 'me u hlapolle ka 100 ml ea PBST 0.05%, kopanya hantle ho fihlela e qhibiliha ka ho feletseng,' me u boloke ho 2-8 ° C.Buffer e lokisitsoeng ea dilution e sebetsa matsatsi a 7.Ho khothalletsoa ho itokisetsa ha ho hlokahala.
3.Tharollo ea ho lemoha 2μg/mL
Nka 48μL ea 0.5 mg/mL Anti-CHO HCP-HRP 'me u hlapolle ka 11,952μL ea 1% BSA ho fumana motsoako oa ho qetela oa 2μg/mL tharollo ea ho lemoha.
4.QC le Tokisetso ea CHOK1 HCP Standards
| Tube Che. | Ea mantlha | Ho tsepamisa maikutlo | Bolumo | 1% BSA | kakaretso ea Bolumo | Qetellong |
| A | Standard | 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 |
Lethathamo: Tokisetso ea QC le Maemo
Mokhoa oa Teko
1.Lokisetsa li-reagents joalo ka ha ho boletsoe ho "Reagent Preparation" ka holimo.
2.Nka 50μL ea litekanyetso, lisampole le QCs (sheba Lethathamo la 3) selibeng se seng le se seng, ebe u eketsa 100μL ea tharollo ea ho lemoha (2μg / mL);Koahela poleiti ka sealer, 'me u behe poleiti ea ELISA holim'a thermomixer.Eketsa ka 500rpm, 25±3℃ bakeng sa lihora tse peli.
3.Fetola microplate ka sekoting ebe u lahla tharollo ea ho roala.Pipette 300μL ea PBST 0.05% ka har'a seliba ka seng ho hlatsoa poleiti ea ELISA le ho lahla tharollo, 'me u phete ho hlatsoa makhetlo a 3.Fetola poleiti holim'a thaole e hloekileng ea pampiri 'me u omelle.
4.Kenya 100μL ea karoloana ea TMB (sheba Lethathamo la 1) selibeng se seng le se seng, tiisa poleiti ea ELISA, 'me u e kene lefifing ho 25±3℃ bakeng sa 15 min.
5.Pipette 100μL ea tharollo ea ho emisa selibeng ka seng.
6.Lekanya ho monya ka bolelele ba 450/650nm ka sebali sa microplate.
7.Sekaseka data ka SoftMax kapa software e lekanang.Etsa moralo o tloaelehileng oa curve ka ho sebelisa mohlala oa tlhophiso ea lintho tse 'nè.
Mohlala oa Mokokotlo o Tloaelehileng
ELA HLOKO: Haeba mahloriso a HCP sampoleng a feta moeli o kaholimo oa curve e tloaelehileng, e hloka ho hlapolloa ka nepo ka buffer ea dilution pele e etsoa tlhahlobo.
LINTLHA TSEO
Tharollo ea ho emisa ke 2M sulfuric acid, ka kopo tšoara ka hloko ho qoba ho phatloha!
