CHO HCP ELISA Kit
Ka whakamahia te tikanga ELISA immunosorbent Kotahi-taahiraa i tenei whakamatautau.Ko nga tauira kei roto CHOK1 HCP ka tauhohe i te wa kotahi me te antibody anti-CHOK1 koati kua tapaina ki te HRP me te paturopi anti-CHOK1 kua pania ki runga i te pereti ELISA, ka mutu ka hanga he matatini hanawiti o te antibody-HCP-waa totoka.Ka taea te tango i te antigen-antibody kaore i herea ma te horoi i te pereti ELISA.Ka taapirihia te tïpako TMB ki te puna mo te tauhohenga rawaka.Ka mutu te whanaketanga o te tae i muri i te taapiri i te otinga aukati, ka panuihia te OD, te uara mimiti ranei o te otinga tauhohenga i te 450/650nm me te panui miihini.Ko te uara OD, te uara mimiti ranei he rite ki te ihirangi HCP i roto i te otinga.Mai i tenei, ka taea te tatau i te kukū o te HCP i roto i te otinga kia rite ki te pihi paerewa.
Taupānga
Ka whakamahia tenei kete ki te kimi ine ine te ihirangi o nga toenga pūmua pūtau kaihautū CHOK1 i roto i nga tauira.
Components
| S/N | Waehanga | Te aro | Nga Tikanga Rokiroki |
| 1 | CHOK1 HCP Paerewa | 0.5mg/mL | ≤–20 ℃ |
| 2 | Anti-CHO HCP-HRP | 0.5mg/mL | ≤–20 ℃, tiaki i te marama |
| 3 | TMB | NA | 2-8 ℃, tiaki i te marama |
| 4 | 20 × PBST 0.05% | NA | 2-8 ℃ |
| 5 | Kati te otinga | NA | RT |
| 6 | Nga kaipatu miihini miihini | NA | RT |
| 7 | BSA | NA | 2-8 ℃ |
| 8 | High adsorption mua-paninga Plates | NA | 2-8 ℃ |
Nga taputapu e hiahiatia ana
| Nga Taonga / Taputapu | Hangahanga | Putumōhio |
| Kaipānui Microplate | Pūrere Molecular | Spectra Max M5, M5e, he rite ranei |
| Whakawerawera | Eppendorf | Eppendorf/5355, he rite ranei |
| Whakaranu Vortex | IKA | MS3 Mamati, he rite ranei |
Te rokiroki me te pumau
1.Te kawe i te -25~-15°C.
2.Ko nga tikanga rokiroki kei te Ripanga 1;Ko nga waahanga 1-2 kei te rongoa ≤–20°C, 5-6 kei te rongoa RT,3,4、7、8 kei te rongoa i te 2-8 ℃;12 marama te wa mana.
Tawhā hua
1.Tairongo: 1ng/mL
2.Awhe rapunga:3- 100ng/mL
3.Tikanga: CV≤ 10% ki roto-whakamatau, CV≤ 15%
4.HCP kapinga: >80%
5.Tauwhāititanga: Ko tenei kete kei te ao katoa na te mea he tauhohenga ki te CHOK1 HCP motuhake mai i te mahi purenga.
Te whakarite reagent
1.PBST 0.05%
Tangohia 15 ml o 20×PBST 0.05%, diluted ki ddH2O, me te hanga ki runga ki te 300 ml.
2.1.0% BSA
Tangohia te 1 karamu o te BSA mai i te pounamu ka waimeha ki te 100 ml o te PBST 0.05%, uru pai kia rewa rawa, ka rongoa ki te 2-8°C.E 7 ra te mana o te waimeha kua oti te whakarite.E taunaki ana kia whakareri ina hiahiatia.
3.Wairewa rapunga 2μg/mL
Tangohia te 48μL o te 0.5 mg/mL Anti-CHO HCP-HRP me te waimeha ki te 11,952μL o te 1% BSA ki te whiwhi i te kukū whakamutunga o te otinga kitenga 2μg/mL.
4.QC me te Whakaritenga o CHOK1 HCP Paerewa
| ngongo Kao. | Taketake | Te aro | Rōrahi | 1%BSA | Rōrahi katoa | Whakamutunga |
| A | Paerewa | 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 |
Ripanga: Whakaritenga o QC me nga Paerewa
Te Mahi Whakatau
1.Whakapaia nga reagents e tohuhia ana i te "Reagent Preparation" i runga ake nei.
2.Tangohia kia 50μL o nga paerewa, tauira me nga QC (tirohia te Ripanga 3) ki ia puna, katahi ka tapirihia te 100μL o te otinga Rapu (2μg/mL);Hipokina te pereti ki te hirikara, ka tuu te pereti ELISA ki runga i te whakamahana mahana.Whakanuia i te 500rpm, 25±3 ℃ mo te 2 haora.
3.Hurihia te pereti moroiti i roto i te totohu ka rukea te otinga paninga.Pipeto 300μL o te PBST 0.05% ki ia puna hei horoi i te pereti ELISA me te whakakore i te otinga, ka whakahoki ano i te horoinga kia toru nga wa.Hurihia te pereti ki runga i te tauera pepa ma, ka patipati kia maroke.
4.Tāpirihia he 100μL o te tïpako TMB (tirohia te Ripanga 1) ki ia puna, hiritia te pereti ELISA, ka whakakororia i roto i te pouri i te 25±3 ℃ mo te 15 min.
5.Pipeto 100μL o te otinga mutu ki ia puna.
6.Te ine i te horonga i te roangaru o te 450/650nm me te panui miihiniiti.
7.Tātarihia te raraunga mā SoftMax, pūmanawa ōrite rānei.Whakatakotoria te ānau paerewa ma te whakamahi i te tauira whakahekenga arorau e wha-tawhā.
Tauira Paerewa Paerewa
FAKATOKANGA: Ki te nui ake te kukū o te HCP i roto i te tauira i te tepe o runga o te ananga paerewa, me tino waimeha ki te parepare waimeha i mua i te whakamatautau.
HE KORERO
Ko te otinga aukati ko te waikawa pungatara 2M, me ata whakahaere kia kore e pakaru!
