Proteinase K (Lyophilized powder)
Nambala ya mphaka: HC4500A
Proteinase K ndi serine protease yokhazikika yokhala ndi gawo lalikulu la gawo lapansi.Imawononga mapuloteni ambiri m'dziko lakwawo ngakhale pamaso pa zotsukira.Umboni wochokera ku maphunziro a kristalo ndi mamolekyu akuwonetsa kuti enzymeyi ndi ya banja la subtilisin lomwe lili ndi malo othandizira othandizira atatu (Asp.39-Wake69- Ser224).Malo odziwika kwambiri a cleavage ndi chomangira cha peptide choyandikana ndi gulu la carboxyl la aliphatic ndi zonunkhira za amino acid okhala ndi magulu otsekedwa a alpha amino.Amagwiritsidwa ntchito kwambiri pakukula kwakemwachindunji.
Zosungirako
2-8 ℃ kusungirako kwakanthawi, -25 ~ -15 ℃ kusungirako nthawi yayitali.Dry ufa udindo -25 ~ -15 ℃ zomveka kwa zaka 3;ufa wa enzyme uyenera kusungunuka mu voliyumu yoyenera.
Kufotokozera
| Maonekedwe | White mpaka woyera amorphous lyophilized ufa |
| Zochita | ≥30 U/mg |
| DNase | Palibe chomwe chapezeka |
| RNase | Palibe chomwe chapezeka |
Katundu
| EC nambala | 3.4.21.64 (Recombinant from Tritirachium album) |
| Kulemera kwa maselo | 29 kDa (SDS-PAGE) |
| Isoelectric point | 7.81 |
| Mulingo wabwino kwambiri wa pH | 7.0-12.0 Mku.1 |
| Kutentha koyenera | 65 ℃ Chithunzi 2 |
| pH kukhazikika | pH 4.5-12.5 (25 ℃, 16h) Chithunzi 3 |
| Kukhazikika kwamafuta | Pansi pa 50 ℃(pH 8.0, 30min) Chithunzi 4 |
| Zoyambitsa | SDS, urea |
| Zoletsa | diisopropyl fluorophosphate;phenylmethylsulfonyl fluoride |
Mapulogalamu
1. Zida zowunikira ma genetic
2. Zida zochotsera ma RNA ndi DNA
3. Kuchotsa zinthu zopanda mapuloteni kuchokera ku minofu, kuwonongeka kwa zonyansa zamapuloteni, monga katemera wa DNA ndikukonzekera heparin.
4. Kukonzekera kwa chromosome DNA ndi pulsed electrophoresis
5. Western blot
6. Enzymatic glycosylated albumin reagents in vitro diagnosis
Kusamalitsa
Valani magalavu oteteza ndi magalasi pamene mukugwiritsa ntchito kapena poyeza sikelo, ndipo sungani mpweya wabwino mukaugwiritsa ntchito.Izi zitha kuyambitsa kusagwirizana ndi khungu komanso kuyabwa kwamaso.Ngati mutakokedwa, zingayambitse zizindikiro za chifuwa kapena mphumu kapena dyspnea.Zitha kuyambitsa kupsa mtima.
Kuyesa
Tanthauzo la unit
Chigawo chimodzi (U) chimatanthauzidwa ngati kuchuluka kwa enzyme yofunikira kuti hydrolyze casein ipange 1 μmol tyrosine pamphindi pamikhalidwe yotsatirayi.
Reagents kukonzekera
Reagent I: 1g mkaka casein anali kusungunuka mu 50ml wa 0.1M sodium mankwala njira (pH 8.0), chofungatira mu 65-70 ℃ madzi 15mins, analimbikitsa ndi kusungunuka, utakhazikika ndi madzi, kusinthidwa ndi sodium hydroxide kuti pH 8.0, ndi yokhazikika voliyumu. 100 ml.
Reagent II: 0.1M trichloroacetic acid, 0.2M sodium acetate, 0.3M acetic acid.
Reagent III: 0.4M Na2CO3yankho.
Reagent IV: Forint reagent kuchepetsedwa ndi madzi oyera kwa 5 nthawi.
Reagent V: enzyme diluent: 0.1M sodium phosphate solution (pH 8.0).
Reagent VI: yankho la tyrosine: 0, 0.005, 0.025, 0.05, 0.075, 0.1, 0.25 umol / ml tyrosine wosungunuka ndi 0.2M HCL.
Ndondomeko
1. 0.5ml ya reagent I imatenthedwa kale mpaka 37 ℃, onjezerani 0.5ml ya njira ya enzyme, sakanizani bwino, ndi kuumitsa pa 37 ℃ kwa 10mins.
2. Onjezani 1 ml ya reagent II kuti muyimitse zomwe zikuchitika, sakanizani bwino, ndikupitiriza kukulitsa kwa 30mins.
3. Centrifugate reaction solution.
4. Tengani 0.5ml supernatant, onjezerani 2.5ml reagent III, 0.5ml reagent IV, sakanizani bwino ndi kuumitsa pa 37 ℃ kwa 30mins.
5. OD660adadziwika ngati OD1;opanda kanthu gulu ulamuliro: 0.5ml reagent V ntchito m'malo enzyme njira kudziwa OD660ndi OD2, ΔOD=OD1-OD2.
6. L-tyrosine yopindika yokhazikika: 0.5mL yosiyana siyana L tyrosine yankho, 2.5mL Reagent III, 0.5mL Reagent IV mu 5mL centrifuge chubu, incubate mu 37 ℃ kwa 30mins, zindikirani kwa OD660pamitundu yosiyanasiyana ya L-tyrosine, kenako adapeza njira yokhazikika Y=kX+b, pomwe Y ndi kuchuluka kwa L-tyrosine, X ndi OD600.
Kuwerengera
2: Chiwerengero chonse cha yankho la mayankho (mL)
0.5: Kuchuluka kwa enzyme solution (mL)
0.5: Kuchuluka kwamadzimadzi komwe kumagwiritsidwa ntchito potsimikiza kwa chromogenic (mL)
10: Nthawi yochitirapo kanthu (mphindi)
Df: Dilution angapo
CKuchuluka kwa enzyme (mg/mL)
Maumboni
1. Wieger U & Hilz H. FEBS Lett.(1972); 23:77.
2. Wieger U & Hilz H. Biochem.Biophys.Res.Commun.(1971); 44:513.
3. Ndi, H.ndi al.,EUR.J. Biochem.( 1975); 56:103–108.
4. Sambrook Jet al., Kuphatikizika kwa mamolekyulu: Buku la Laboratory, kope lachiwiri, Cold Spring HarborLaboratory Press, Cold Spring Harbor (1989).
Mkuyu. 2 Kutentha kwakukulu
Zomwe zimachitika mu 20 mM K-phosphate buffer pH 8.0.Kuchuluka kwa enzyme: 1 mg/mL
Chithunzi 3 pH Kukhazikika
25 ℃, 16 h-mankhwala ndi 50mM buffer yankho: pH 4.5-5.5, Acetate;pH 6.0-8.0, Na-phosphate;pH 8.0-9.0, Tris-Mtengo wa HCL.pH 9.0-12.5, Glycine-NaOH.Kuchuluka kwa enzyme: 1 mg/mL
Chithunzi cha 4 Thermal bata
30 min-mankhwala ndi 50mM Tris-HCL buffer, pH 8.0.Kuchuluka kwa enzyme: 1 mg/mL
Chithunzi 5 Kusungirako khazikikaty at 25 ℃
