Proteinase K (Amazi)
Injangwe No: HC4502A
Proteinase K ni proteine ihamye hamwe na substrate yagutse.Itesha agaciro poroteyine nyinshi muri kavukire ndetse no imbere yimyenda.Ibimenyetso bivuye mubushakashatsi bwa kristu na molekuline byerekana ko enzyme ari iyumuryango wa subtilisin hamwe na site ikora catalitike triad (Asp39-Yiwe69- Ser224).Ikibanza cyiganjemo clavage ni peptide ihuza peptide yegeranye na carboxyl groupe ya aliphatic na aromatic amino acide hamwe na alpha amino yahagaritswe.Bikunze gukoreshwa mugariumwihariko.
Ibisobanuro
| Kugaragara | Ibara ritagira ibara ryijimye |
| Igikorwa | 00800 U / ml |
| Intungamubiri za poroteyine | ≥20 mg / ml |
| DNase | Nta n'umwe wamenyekanye |
| RNase | Nta n'umwe wamenyekanye |
Ububiko
Ubike ku bushyuhe bwa 2-8 ℃.
Ibyiza
| Numero ya EC | 3.4.21.64 (R.ecombinant yo muri alubumu ya Tritirachium) |
| Uburemere bwa molekile | 29 kDa (SDS-URUPAPURO) |
| Ingingo ya Isoelectric | 7.81 |
| PH nziza | 7.0-12.0 Igishusho.1 |
| Ubushyuhe bwiza | 65 ℃ Igishusho.2 |
| pH ituze | pH 4.5-12.5 (25 ℃, 16 h) Igishusho.3 |
| Ubushyuhe bukabije | Munsi ya 50 ℃ (pH 8.0, iminota 30) Igishusho.4 |
| Abakangurambaga | SDS, urea |
| Inhibitor | Diisopropyl fluorophosphate;fenilmethylsulfonyl fluoride |
Porogaramu
1. Ibikoresho byo gusuzuma
2. Ibikoresho byo gukuramo RNA na ADN
3. Gukuramo ibice bitari poroteyine biva mu ngingo, kwangirika kwa poroteyine, nkinkingo za ADN no gutegura heparin
4. Gutegura ADN ya chromosome na pulsed electrophorei
5. Iburengerazuba
6. Enzymatique glycosylated albumin reagents mugupima vitro
Kwirinda
Wambare uturindantoki two kurinda hamwe na gogles mugihe ukoresha cyangwa upima, kandi ukomeze guhumeka neza nyuma yo kubikoresha.Iki gicuruzwa gishobora gutera uruhu rwa allergique no kurwara amaso.Niba ihumeka, irashobora gutera allergie cyangwa ibimenyetso bya asima cyangwa dyspnea.Birashobora gutera uburakari.
Suzuma
Igisobanuro cyibice
Igice kimwe (U) gisobanurwa nkubunini bwa enzyme isabwa kuri hydrolyze casein kugirango itange 1 μ mol tyrosine kumunota mubihe bikurikira.
Reagents kwitegura
Reagent I: 1g amata yamata yashongeshejwe muri 50ml yumuti wa 0.1M sodium fosifate (pH 8.0), yashizwemo mumazi 65-70 ℃ mumazi 15mins, akayungurura akayashonga, akonjeshwa namazi, agahindurwa na hydroxide ya sodium kuri pH8.0, hanyuma agashyirwaho ingano 100ml.
Reagent II: Igisubizo cya TCA: 0.1M acide trichloroacetic, 0.2M sodium acetate, 0.3M acide acetike.
Reagent III: 0.4M Na2CO3igisubizo.
Reagent IV: Forint reagent ivanze namazi meza inshuro 5.
Reagent V: Enzyme diluent: 0.1M sodium fosifate yumuti (pH 8.0).
Reagent VI: Umuti wa Tyrosine: 0, 0.005, 0.025, 0.05, 0.075, 0.1, 0,25 umol / ml tirozine yashonga hamwe na 0.2M HCl.
Inzira
1. 0.5ml ya reagent Nashyutswe mbere kuri 37 ℃, ongeramo 0.5ml yumuti wa enzyme, vanga neza, kandi ube kuri 37 ℃ kuri 10mins.
2. Ongeramo 1ml ya reagent II kugirango uhagarike reaction, vanga neza, kandi ukomeze incubation kuri 30mins.
3. Centrifugate igisubizo cyibisubizo.
4. Fata 0.5ml ndengakamere, ongeramo 2.5ml reagent III, 0.5ml reagent IV, vanga neza hanyuma ushire kuri 37 ℃ kuri 30mins.
5. OD660yagenwe nka OD1;itsinda rishinzwe kugenzura ubusa: 0.5ml reagent V ikoreshwa mugusimbuza enzyme igisubizo kugirango umenye OD660nka OD2, ΔOD = OD1-OD2.
6660kubintu bitandukanye bya L-tyrosine, hanyuma ibona umurongo usanzwe Y = kX + b, aho Y ni L-tyrosine yibanze, X ni OD600.
Kubara
2: Umubare wuzuye wibisubizo (mL)
0.5: Ingano yumuti wa enzyme (mL)
0.5: Igipimo cyamazi yakoreshejwe mugukoresha chromogenic (mL)
10: Igihe cyo kubyitwaramo (min)
Df: Gukoresha byinshi
C: Enzyme yibanze (mg / mL)
Reba
1. Wieger U & Hilz H. FEBS Mureke.(1972);23:77.
2. Wieger U & Hilz H. Biochem.Biophys.Res.Umuganda.(1971);44: 513.
3. Hilz, H.n'abandi.,Eur.J. Biochem.(1975);56: 103-108.
4. Sambrook J.et al., Molecular Cloning: Igitabo cya Laboratoire, integuro ya 2, Itangazamakuru rya Cold Spring Harbour Laboratoire, Cold Spring Harbour (1989).
Imibare
Igishushanyo. 1 Ibyiza pH
100mM igisubizo cya buffer: pH6.0-8.0, Na-fosifate;pH8.0- 9.0, Tris-HCl;pH9.0-12.5, Glycine-NaOH.Ubunini bwa Enzyme: 1mg / mL
Igishushanyo cya 2 Ubushyuhe bwiza
Igisubizo muri 20mM K-fosiferi buffer pH 8.0.Enzyme yibanze: 1mg / mL
Igishushanyo cya 3 pH Igihagararo
25 ℃, 16 h-kuvura hamwe na 50mM igisubizo cya buffer: pH 4.5-5.5, Acetate;pH 6.0-8.0, Na-fosifate;pH 8.0-9.0, Tris-HCl.pH 9.0-12.5, Glycine-NaOH.Enzyme yibanze: 1mg / mL
Igishushanyo cya 4 Ubushyuhe ituze
30 min-kuvura hamwe na 50mM Tris-HCl buffer, pH 8.0.Enzyme yibanze: 1mg / mL
Igishushanyo cya 5 Ububiko stability at 25 ℃
