CHO HCP ELISA Kit
Ana amfani da hanyar ELISA immunosorbent mataki ɗaya a cikin wannan binciken.Samfuran da ke ɗauke da CHOK1 HCP a lokaci guda suna amsawa tare da HRP mai lakabin goat anti-CHOK1 antibody da anti-CHOK1 antibody wanda aka lulluɓe akan farantin ELISA, a ƙarshe ya samar da hadaddun sanwici na antibody mai ƙarfi mai ƙarfi-HCP.Ana iya cire antigen-antigen da ba a ɗaure ba ta hanyar wanke farantin ELISA.Ana ƙara ma'aunin TMB zuwa rijiyar don isasshiyar amsawa.Ana dakatar da haɓakar launi bayan ƙara maganin tasha, kuma ana karanta OD ko ƙimar ƙimar maganin amsawa a 450/650nm tare da mai karanta microplate.Ƙimar OD ko ƙimar abin sha ya yi daidai da abun ciki na HCP a cikin maganin.Daga wannan, ana iya ƙididdige ƙaddamarwar HCP a cikin bayani bisa ga daidaitattun ma'auni.
Aikace-aikace
Ana amfani da wannan kit ɗin don gano ainihin abun ciki na CHOK1 ragowar furotin tantanin halitta a cikin samfurori.
Cmasu kai hari
| S/N | Bangaren | Hankali | Yanayin Ajiya |
| 1 | Farashin CHOK1 HCP | 0.5mg/ml | ≤-20℃ |
| 2 | Anti-CHO HCP-HRP | 0.5mg/ml | ≤–20℃, kariya daga haske |
| 3 | TMB | NA | 2-8 ℃, kariya daga haske |
| 4 | 20 × PBST 0.05% | NA | 2-8 ℃ |
| 5 | Tsaida mafita | NA | RT |
| 6 | Microplate sealers | NA | RT |
| 7 | BSA | NA | 2-8 ℃ |
| 8 | Babban adsorption pre-shafi faranti | NA | 2-8 ℃ |
Ana Bukatar Kayan aiki
| Kayayyakin Kayayyaki/Kayan Kayayyaki | Kerawa | Katalogi |
| Mai karanta Microplate | Na'urorin Kwayoyin Halitta | Spectra Max M5, M5e, ko makamancin haka |
| Thermomixer | Eppendorf | Eppendorf/5355, ko makamancin haka |
| Mai haɗa Vortex | IKA | MS3 Digital, ko makamancin haka |
Adana da kwanciyar hankali
1.Kai a -25 ~ -15°C.
2.Yanayin ajiya suna kamar yadda aka nuna a cikin Table 1;aka adana 1-2 ≤-20 ° C, 5-6 ana adana RT,3,4,7,8 ana adana a 2-8 ℃;lokacin aiki shine watanni 12.
Siffofin samfur
1.Hankali: 1ng/ml
2.Kewayon ganowa: 3-100ng/ml
3.Matsakaicin: Intra-assay CV≤ 10%, inter-assay CV≤ 15%
4.HCP:> 80%
5.Ƙayyadaddun bayanai: Wannan kit ɗin na duniya ne yayin da yake amsawa ta musamman tare da CHOK1 HCP mai zaman kansa daga tsarin tsarkakewa.
Reagent shiri
1.PBST 0.05%
Ɗauki 15 ml na 20×PBST 0.05%, diluted a ddH2O, kuma an yi har zuwa 300 ml.
2.1.0% BSA
Ɗauki 1g na BSA daga kwalban kuma a tsoma a cikin 100 ml na PBST 0.05%, gauraya sosai har sai an narkar da shi gaba daya, kuma adana a 2-8 ° C.Shirye-shiryen dilution buffer yana aiki na kwanaki 7.Ana bada shawara don shirya yadda ake bukata.
3.Maganin ganowa 2μg/ml
Ɗauki 48μL na 0.5 mg/mL Anti-CHO HCP-HRP kuma tsarma a cikin 11,952μL na 1% BSA don samun taro na ƙarshe na 2μg/mL mafita ga ganowa.
4.QC da Shiri na CHOK1 HCP Standards
| Tube A'a. | Na asali | Hankali | Ƙarar | 1% BSA | jimlar girma | Karshe |
| A | Daidaitawa | 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 |
Tebur: Shirye-shiryen QC da Matsayi
Hanyar tantancewa
1.Shirya reagents kamar yadda aka nuna a cikin "Shirin Reagent" a sama.
2.Ɗauki 50μL na ma'auni, samfurori da QCs (koma zuwa Table 3) a cikin kowace rijiya, sannan ƙara 100μL na Maganin Gano (2μg / ml);Rufe farantin tare da sealer, kuma sanya farantin ELISA akan thermomixer.Incubate a 500rpm, 25± 3℃ na 2 hours.
3.Juya microplate a cikin kwatami kuma jefar da maganin rufewa.Pipette 300μL na PBST 0.05% a cikin kowace rijiya don wanke farantin ELISA kuma zubar da maganin, kuma maimaita wanke sau 3.Juya farantin a kan tawul ɗin takarda mai tsabta kuma a bushe.
4.Ƙara 100μL na TMB substrate (duba Table 1) zuwa kowace rijiya, rufe farantin ELISA, da kuma sanyawa a cikin duhu a 25 ± 3 ℃ na 15 min.
5.Pipette 100μL na maganin tasha a cikin kowace rijiya.
6.Auna sha a tsawon 450/650nm tare da mai karanta microplate.
7.Yi nazarin bayanai ta SoftMax ko software makamancin haka.Yi ƙirƙira daidaitaccen lanƙwasa ta amfani da ƙirar juzu'i mai ma'ana huɗu.
Misalin Lanƙwasa Madaidaici
NOTE: Idan maida hankali na HCP a cikin samfurin ya wuce iyakar babba na daidaitaccen lanƙwasa, yana buƙatar a shafe shi da kyau tare da buffer dilution kafin gwaji.
BAYANI
Maganin tsayawa shine 2M sulfuric acid, da fatan za a rike da kulawa don guje wa splashing!
