Ultra Nuclease GMP-ọkwa
Mpempe nwamba: HC2016A
UltraNuclease GMP-ọkwa ka egosipụtara ma sachaa ya na Escherichia coli (E.coli) site n'usoro mkpụrụ ndụ ihe nketa na kwadebere n'okpuru gburugburu GMP.Ọ nwere ike ibelata viscosity nke cell supernatant na cell lysate na nyocha sayensị, na-abawanye nrụpụta protein dị ọcha ma kwalite nyocha ọrụ protein.Ngwaahịa ahụ nwekwara ike ibelata ihe fọdụrụ n'ime acid nucleic acid na pg-grade, na-eme ka arụmọrụ na nchekwa nke ngwaahịa ngwa ngwa gụnyere ọcha nje, mmepụta ọgwụ mgbochi, na mmepụta protein/polysaccharide ọgwụ.E wezụga nke ahụ, a pụkwara itinye ngwaahịa a iji gbochie nchikota nke mkpụrụ ndụ mononuclear ọbara mmadụ (PBMC) na ọgwụgwọ cell na mmepe ọgwụ mgbochi.
A na-enye UltraNuclease n'ụdị reagent igba ogwu, nke dị elu na nchekwa (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), na ọdịdị nke mmiri na-enweghị agba, nke na-egbuke egbuke.Emepụtara ngwaahịa a site na usoro GMP chọrọ ma nye ya n'ụdị mmiri mmiri.
Ngwa
UltraNuclease GMP-ọkwa (250 U/μL)
Ọnọdụ Nchekwa
A na-ebufe ngwaahịa a na ice kpọrọ nkụ ma nwee ike ịchekwa ya na -25 ℃ ~ -15 Celsius maka afọ abụọ.
Ọ bụrụ na emepere ngwaahịa ahụ ma chekwaa ya na 4 ℃ ihe karịrị otu izu, anyị na-akwado nzachangwaahịa ahụ iji gbochie mmetọ microbial.
Nkọwapụta
| Onye ọbịa okwu | Recombinant E. coli nwere mkpụrụ ndụ ihe nketa UltraNuclease |
| Ibu molekụla | 26,5kDa |
| ebe soelectric | 6.85 |
| Ịdị ọcha | ≥99% (SDS-PAGE) |
| Nchekwa nchekwa | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin |
|
Nkọwa nkeji | Nkọwa nke otu ngalaba ọrụ (U) bụ ọnụọgụ enzyme ejigbanwee uru nnabata nke ΔA260 site na 1.0 n'ime nkeji 30 na 2. 625 mlusoro mmeghachi omume na 37 ℃ na pH nke 8.0 (dakọtara na mgbaze zuru oke37 μg salmon spam DNA n'ime oligonucleotides). |
Ntuziaka
1. Ihe atụ Nchịkọta
Selụ agbakwunyere: wepụ ihe ọkara, jiri PBS sachaa mkpụrụ ndụ, ma wepụ ihe dị elu.
Mkpụrụ ndụ nkwusioru: na-anakọta mkpụrụ ndụ site na centrifugation, sachaa mkpụrụ ndụ na PBS, centrifuge na 6,000rpm maka 10 min, nakọta pellet.
Escherichia coli: nakọta nje bacteria site na centrifugation, saa otu ugboro na PBS, centrifuge na 8,000rpm maka 5 min, ma nakọta pellet.
2. Ọgwụgwọ sample
Na-emeso pellets ndị anakọtara na ihe nchekwa lysis na oke nke uka (g) na olu (mL) 1: (10-20), ma ọ bụ site na usoro nhazi ma ọ bụ kemịkalụ na ice ma ọ bụ na okpomọkụ (1g nke pellet cell nwere ihe dịka
109 sel).
3. Ọgwụgwọ Enzyme
Tinye 1-5mM MgCl na usoro mmeghachi omume ma mezie pH na 8-9.
Tinye UltraNuclease dị ka nha nke 250 nkeji iji gbarie 1 g nke pellet cell, tinye na 37 ℃ maka ihe karịrị nkeji 30.Biko rụtụ aka n'ụdị "Oge mmeghachi omume akwadoro" ka ịhọrọ yaoge ọgwụgwọ.
4. Supernatant Nchịkọta
Centrifuge na 12,000 rpm maka nkeji 30 wee kpokọta ihe dị elu.
Rịba ama: Ọ bụrụ na ngwọta bụ acidic ma ọ bụ alkaline, ma ọ bụ nwere nnukwu nnu, ncha, ma ọ bụdenaturants, biko welie usoro onunu ogwu ma ọ bụ gbatịa oge ọgwụgwọ ya.
Mmeghachi omume akwadoro conditions
| Oke | Ọnọdụ kacha mma | Ọnọdụ dị irè |
| Mg²+Ntụkwasị obi | 1-5 mm | 1-10 mm |
| pH | 8-9 | 6-10 |
| Okpomọkụ | 37 ℃ | 0-42 ℃ |
| Ntinye uche DTT | 0-100 mm | > 0 mm |
| Mkpokọta Mercaptoethanol | 0-100 mm | > 0 mm |
| Nchịkọta Monovalent Cation | 0-20 mm | 0-150 mm |
| Phosphate lon itinye uche | 0-10 mm | 0-100 mm |
Akwadoro Mmeghachi omume Oge (37 ℃, 2 mM Mg²+, pH 8.0)
| Ọnụ ego UltraNuclease (Ntụkwasị Obi Ikpeazụ) | Oge mmeghachi omume |
| 0.25 U/ml | >10 h |
| 2.5 U/ml | > 4h |
| 25 U/ml | 30 min |
ndetu:
Biko yiri PPE dị mkpa, uwe ụlọ nyocha na uwe aka, iji hụ na ahụike na nchekwa gị!
