CHO HCP ELISA Kit
A na-eji usoro ELISA immunosorbent otu nzọụkwụ mee ihe na nyocha a.Nlereanya nwere CHOK1 HCP na-emeghachi omume n'otu oge na ewu mgbochi CHOK1 mgbochi HRP na mgbochi CHOK1 ekpuchiri n'efere ELISA, n'ikpeazụ na-etolite ogige Sanwichi nke mgbochi siri ike-phase-HCP-labeled antibody.Enwere ike iwepụ antigen-antibody na-ekekọta site na ịsacha efere ELISA.A na-agbakwunye mkpụrụ TMB n'olulu mmiri maka mmeghachi omume zuru oke.Akwụsị mmepe nke agba ahụ mgbe agbakwunyere nkwụsị nkwụsị, na OD ma ọ bụ uru nnabata nke ngwọta mmeghachi omume na 450/650nm na-agụ ya na onye na-agụ microplate.Uru OD ma ọ bụ uru nnabata dabara na ọdịnaya HCP dị na ngwọta.Site na nke a, enwere ike gbakọọ mkpokọta HCP na ngwọta dịka usoro ọkọlọtọ.
Ngwa
A na-eji ngwa a iji chọpụta ọnụọgụgụ ọdịnaya nke CHOK1 foduru protein cell host na nlele.
Cndị ọchịchị
| S/N | Akụkụ | Ntinye uche | Ọnọdụ Nchekwa |
| 1 | Ọkọlọtọ CHOK1 HCP | 0.5mg/mL | ≤-20℃ |
| 2 | Mgbochi CHO HCP-HRP | 0.5mg/mL | ≤–20℃, chebe pụọ na ìhè |
| 3 | TMB | NA | 2-8 ℃, chebe site na ìhè |
| 4 | 20 × PBST 0.05% | NA | 2-8 ℃ |
| 5 | Kwụsị ngwọta | NA | RT |
| 6 | Microplate sealers | NA | RT |
| 7 | BSA | NA | 2-8 ℃ |
| 8 | Akwa adsorption tupu mkpuchi efere | NA | 2-8 ℃ |
Ngwa achọrọ
| Ngwa / Akụrụngwa | Mepụta | Katalọgụ |
| Onye na-agụ Microplate | Ngwaọrụ molekụla | Spectra Max M5, M5e, ma ọ bụ dakọtara |
| Thermomixer | Eppendorf | Eppendorf/5355, ma ọ bụ otu |
| Ngwakọta Vortex | IKA | MS3 Digital, ma ọ bụ dakọtara |
Nchekwa na nkwụsi ike
1.Ụgbọ njem na -25 ~ -15 Celsius.
2.Ọnọdụ nchekwa dị ka egosiri na Tebụl 1;A na-echekwa akụkụ 1-2 ≤-20 ° C, 5-6 na-echekwa RT,3,4,7,8 na-echekwara na 2-8 ℃;oge nkwado bụ ọnwa iri na abụọ.
Ngwa ngwaahịa
1.Mmetụta uche: 1ng/ml
2.Oke nchọpụta: 3-100ng/ml
3.Nkenke: Intra-assay CV≤ 10%, inter-assay CV≤ 15%
4.Mkpuchi HCP:> 80%
5.Nkọwapụta: Ngwa a zuru ụwa ọnụ ka ọ na-emeghachi omume na CHOK1 HCP na-adabereghị na usoro nchacha.
Nkwadebe reagent
1.PBST 0.05%
Were 15 ml nke 20 × PBST 0.05%, gbazere na ddH2O, ma mee ihe ruru 300 ml.
2.1.0% BSA
Were 1g nke BSA site na karama ma gbanye na 100 ml nke PBST 0.05%, gwakọta nke ọma ruo mgbe agbazere kpamkpam, ma chekwaa na 2-8 ° C.Ihe nchekwa dilution akwadoro dị irè maka ụbọchị 7.A na-atụ aro ka ị kwadebe dị ka ọ dị mkpa.
3.Nchọpụta nchọpụta 2μg/ml
Were 48μL nke 0.5 mg/mL Anti-CHO HCP-HRP na itughari na 11,952μL nke 1% BSA iji nweta ntinye ikpeazụ nke 2μg/mL ngwọta nchọpụta.
4.QC na Nkwadebe nke ụkpụrụ CHOK1 HCP
| Tube Mba. | Nke mbụ | Ntinye uche | Olu | 1% BSA | mkpokọta Olu | Ikpeazụ |
| A | Ọkọlọtọ | 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 |
Tebụl: Nkwadebe nke QC na ụkpụrụ
Usoro nyocha
1.Kwadebe reagents dị ka egosiri na "Reagent Preparation" n'elu.
2.Were 50μL nke ụkpụrụ, ihe atụ na QCs (na-ezo aka na Tebụl 3) n'ime olulu mmiri ọ bụla, wee tinye 100μL nke ngwọta nchọpụta (2μg / mL);Na-ekpuchi efere na sealer, ma tinye efere ELISA na thermomixer.Incubate na 500rpm, 25± 3℃ maka awa 2.
3.Tụgharịa microplate na sink ma tụfuo ngwọta mkpuchi.Pipette 300μL nke PBST 0.05% n'ime olulu mmiri ọ bụla iji sachaa efere ELISA wee tụfuo ihe ngwọta, ma megharịa ịsa ahụ ugboro 3.Tụgharịa efere ahụ na akwa nhicha akwụkwọ dị ọcha wee kpoo ya.
4.Tinye 100μL nke TMB mkpụrụ (lee Tebụl 1) na olulu mmiri ọ bụla, mechie efere ELISA, ma tinye n'ọchịchịrị na 25± 3℃ maka 15 min.
5.Pipette 100μL ngwọta nkwụsị n'ime olulu mmiri ọ bụla.
6.Tụọ nnabata na ogologo wavelength nke 450/650nm na onye na-agụ microplate.
7.Nyochaa data site na SoftMax ma ọ bụ ngwa ngwa dakọtara.Kpaa nkata ọkọlọtọ site na iji ihe nlere mgbagha ihe mgbagha anọ.
Ọkọlọtọ Curve Ihe Nlereanya
IHE: Ọ bụrụ na ntinye nke HCP dị na nlele ahụ gafere oke elu nke usoro ọkọlọtọ, ọ kwesịrị ka etinye ya nke ọma na ihe nchekwa dilution tupu nnwale.
NKWUO
Ngwọta nkwụsị bụ 2M sulfuric acid, biko jikwaa nke ọma ka ị ghara ịgbasa!
