Phosphatase Alkaline (ALP)
Nkọwa
Alkaline Phosphatase sitere na nje E. coli recombinant nke na-ebu mkpụrụ ndụ TAB5.Enzyme ahụ na-akwalite dephosphorylation nke 5' na 3' nsọtụ DNA na RNA phosphomonoesters.Ọzọkwa, ọ hydrolyses ribose, yana deoxyribonucleoside triphosphates (NTPs na dNTPs).TAB5 Alkaline Phosphatase na-arụ ọrụ na 5 ′ na-apụta, 5′ agbapụta na njedebe na-enweghị isi.Enwere ike iji Phosphatase n'ọtụtụ ngwa usoro ihe ọmụmụ ihe, dị ka cloning ma ọ bụ akara njedebe nyocha iji wepụ nsọtụ phosphorylated nke DNA ma ọ bụ RNA.N'ime nyocha nke cloning, dephosphorylation na-egbochi DNA plasmid linearized site na njide onwe onye.Ọ nwekwara ike weda dNTP enweghị njikọ na nzaghachi PCR iji kwado ndebiri maka usoro DNA.Enzyme ahụ kpamkpam na enweghị ike ịgbagha ya site na ikpo ọkụ na 70 Celsius C maka nkeji 5, si otú a na-eme ka mwepụ nke phosphatase tupu ligation ma ọ bụ njedebe na-enweghị isi.
Ojiji
1.Alkaline phosphatase jikọtara ya na protein (ọgwụ mgbochi ọrịa, streptavidin wdg,) nwere ike ịmata kpọmkwem mkpụrụ ndụ lekwasịrị anya, a pụkwara iji ya mee ihe na ELISA, WB na nchọpụta histochemical;
2.Alkaline phosphatase nwere ike iji dephosphorize 5 '-terminal nke DNA ma ọ bụ RNA iji gbochie njikọ onwe;
3. DNA ma ọ bụ RNA dephosphorylated dị n'elu nwere ike ịkpọ ya site na redio phosphates (site na T4 poly-nucleotide kinase)
Ọdịdị Chemical
Nkọwapụta
Nnwale ihe | Nkọwapụta |
Enzyme ọrụ | 5U/μL |
Ọrụ Endonuclease | Achọpụtaghị |
Ọrụ Exonuuclease | Achọpụtaghị |
Ọrụ Nicking | Achọpụtaghị |
Ọrụ RNase | Achọpụtaghị |
E.coli DNA | ≤1 oyiri/5U |
Endotoxin | Nnwale LAL, ≤10EU/mg |
Ịdị ọcha | ≥95% |
Ụgbọ njem na nchekwa
Ụgbọ njem:Ambient
Nchekwa:Na-echekwa na 2-8°C
Nnwale akwadoroNdụ:Afọ 2