CHO HCP ELISA Kit
Hoʻohana ʻia kahi ʻano hana immunosorbent ELISA hoʻokahi i kēia hoʻokolohua.Hoʻopili like nā laʻana i loko o ka CHOK1 HCP me ka HRP-labeled goat anti-CHOK1 antibody a me anti-CHOK1 antibody i uhi ʻia ma ka pā ELISA, ma hope o ka hana ʻana i kahi paʻa sandwich o ka antibody-HCP-labeled antibody.Hiki ke wehe ʻia ka antigen-antibody i hoʻopaʻa ʻole ʻia ma ka holoi ʻana i ka pā ELISA.Hoʻohui ʻia ka substrate TMB i ka pūnāwai no ka pane kūpono.Hoʻopau ʻia ka hoʻomohala kala ma hope o ka hoʻohui ʻana i ka solution stop, a heluhelu ʻia ka OD a i ʻole ka waiwai absorbance o ka hopena hopena ma 450/650nm me kahi mea heluhelu microplate.ʻO ka waiwai OD a i ʻole ka waiwai absorbance e like me ka HCP maʻiʻo i loko o ka hoʻonā.Mai kēia, hiki ke helu ʻia ka ʻike HCP i ka hoʻonā e like me ka pihi maʻamau.
Palapala noi
Hoʻohana ʻia kēia kit e ʻike nui i ka ʻike o nā koena protein cell host CHOK1 i nā laʻana.
Cnā mea hoʻokūkū
S/N | ʻāpana | Hoʻopaʻa | Nā Kūlana Waihona |
1 | CHOK1 HCP Kūlana | 0.5mg/mL | ≤–20 ℃ |
2 | Anti-CHO HCP-HRP | 0.5mg/mL | ≤–20 ℃, pale mai ka malamalama |
3 | TMB | NA | 2-8 ℃, pale mai ka malamalama |
4 | 20 × PBST 0.05% | NA | 2-8 ℃ |
5 | Hoʻopau i ka hoʻonā | NA | RT |
6 | ʻO nā mea hōʻailona microplate | NA | RT |
7 | BSA | NA | 2-8 ℃ |
8 | High adsorption pre-coating Pā | NA | 2-8 ℃ |
Pono nā lako
Mea hoopau / Lako | Hana ʻia | Heluhelu |
Heluhelu microplate | Nā Mea Molekala | Spectra Max M5, M5e, a i ʻole like |
ʻO ka thermomixer | ʻEppendorf | Eppendorf/5355, a i ʻole like |
Mea hui puana | IKA | MS3 Digital, a i ʻole like |
Ka mālama ʻana a me ke kūpaʻa
1.Lawe ʻia ma -25~-15°C.
2.ʻO nā kūlana mālama e like me ia i hōʻike ʻia ma ka Papa 1;mālama ʻia nā ʻāpana 1-2 ≤–20°C, mālama ʻia 5-6 RT,3,4、7、8 mālama ʻia ma 2-8 ℃;ʻO ka manawa kūpono he 12 mahina.
Nā palena huahana
1.ʻIke: 1ng/mL
2.ʻIke ʻia: 3- 100ng/mL
3.Ka pololei: Intra-assay CV≤ 10%, inter-assay CV≤ 15%
4.HCP uhi: >80%
5.Mea kiko'ī: He mea āpau kēia kit no ka mea e hoʻopili pono ʻia me CHOK1 HCP kūʻokoʻa i ke kaʻina hoʻomaʻemaʻe.
Ka hoʻomākaukau ʻana i ka reagent
1.PBST 0.05%
E lawe i 15 ml o 20×PBST 0.05%, hoʻoheheʻe ʻia i ka ddH2O, a hiki i ka 300 ml.
2.1.0% BSA
E lawe i 1g o ka BSA mai ka omole a hoʻoheheʻe ʻia i loko o 100 ml o PBST 0.05%, hui maikaʻi a hiki i ka hoʻoheheʻe ʻana, a mālama i ka 2-8 ° C.Pono ka dilution buffer i hoʻomākaukau ʻia no 7 mau lā.Paipai ʻia e hoʻomākaukau e like me ka mea e pono ai.
3.Hoʻonā ʻike 2μg/mL
E lawe i 48μL o 0.5 mg/mL Anti-CHO HCP-HRP a hoʻoheheʻe ʻia ma 11,952μL o 1% BSA e loaʻa ai ka hopena hope loa o 2μg/mL ʻike.
4.QC a me ka hoʻomākaukau ʻana i nā Kūlana CHOK1 HCP
Paipu ʻAʻole. | Kumu | Hoʻopaʻa | Volume | 1%BSA | huina Volume | hope loa |
A | Kūlana | 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 |
Papa: Hoʻomākaukau i ka QC a me nā Kūlana
Kaʻina hana hoʻokolohua
1.E hoʻomākaukau i nā reagents e like me ka mea i hōʻike ʻia ma ka "Reagent Preparation" ma luna.
2.E lawe i 50μL o nā maʻamau, nā laʻana a me nā QC (e nānā i ka Papa 3) i loko o kēlā me kēia pūnāwai, a laila e hoʻohui i 100μL o ka Detection solution (2μg/mL);E uhi i ka pā me ka mea sila, a kau i ka pā ELISA ma luna o ka thermomixer.Hoʻomoʻa ma 500rpm, 25±3 ℃ no 2 mau hola.
3.E hoʻohuli i ka microplate i loko o ka ipuhao a hoʻolei i ka ʻaha hoʻouhi.Pipette 300μL o PBST 0.05% i loko o kēlā me kēia pūnāwai e holoi i ka pā ELISA a hoʻolei i ka hopena, a hana hou i ka holoi ʻana 3 manawa.E hoʻohuli i ka pā ma kahi kāwele pepa maʻemaʻe a maloʻo.
4.E hoʻohui i 100μL o TMB substrate (e nānā i ka Papa 1) i kēlā me kēia punawai, e sila i ka pā ELISA, a hoʻomoʻa i ka pōʻeleʻele ma 25±3 ℃ no 15 min.
5.Pipette 100μL o ka solution stop i loko o kēlā me kēia pūnāwai.
6.E ana i ka hawewe ma ka nalu nalu o 450/650nm me ka mea heluhelu microplate.
7.E noʻonoʻo i ka ʻikepili e SoftMax a i ʻole nā polokalamu like.E hoʻolālā i ka pihi maʻamau ma o ka hoʻohana ʻana i ke kumu hoʻohālikelike logistic regression ʻehā.
Laʻana Pipi maʻamau
NĀ MEA: Inā ʻoi aku ka nui o ka HCP i loko o ka hāpana ma mua o ka palena kiʻekiʻe o ka pihi maʻamau, pono e hoʻoheheʻe ʻia me ka dilution buffer ma mua o ka hoʻāʻo ʻana.
OLELO HOOLAHA
ʻO ka hopena hoʻomaha ʻo 2M sulfuric acid, e ʻoluʻolu e mālama me ka mālama pono e pale aku i ka splashing!