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Imikhiqizo
I-Proteinase K NGS (impuphu) HC4507A Isithombe Esifakiwe
  • Iphrotheni K NGS (powder) HC4507A
  • Iphrotheni K NGS (powder) HC4507A

I-Proteinase K NGS (impuphu)


Inombolo yekati: HC4507A

Iphakheji: 1g/10g/100g/500g

 Mahhala DNase, RNase, Nickase

Umsebenzi: ≥40 U/mg

Izinsalela ze-Nucleic Acid: ≤ 5 pg/mg

Umthwalo wezinto eziphilayo: ≤ 50 CFU/g

Impilo yeshelufu iminyaka emi-3

Ezokuthutha ekamelweni lokushisa

Umthamo weqoqo elilodwa 30kg

 

  • sns01
  • sns03
  • sns04
  • sns05

Incazelo Yomkhiqizo

Imininingwane yomkhiqizo

Idatha

Inombolo yekati: HC4507A

I-NGS Protease K iyi-serine protease ezinzile enomsebenzi we-enzyme ephezulu kanye nokucaciswa kwe-substrate ebanzi.I-enzyme ikhetha ukubola amabhondi e-ester namabhondi e-peptide eduze kwe-C-terminal yama-hydrophobic amino acid, ama-amino acid anesulfure nama-amino acid anephunga elimnandi.Ngakho-ke, kuvame ukusetshenziselwa ukwehlisa amaprotheni abe ama-peptide amafushane.I-NGS Protease K iyi-serine protease ejwayelekile ene-Asp39-Wakhe69-Ser224i-catalytic triad ehlukile kuma-serine proteases, futhi isikhungo se-catalytic sizungezwe yi-tow Ca2+izindawo ezibophayo zokuzinzisa, ukugcina umsebenzi ophezulu we-enzyme ngaphansi kobubanzi obubanzi bezimo.

  • Okwedlule:
  • Olandelayo:

    • Ukucaciswa

    Ukubukeka

    Impushana ye-amorphous emhlophe ukuya kokumhlophe, i-lyophilized

    Umsebenzi othize

    ≥40U/mg okuqinile

    DNase

    Akukho okutholiwe

    RNase

    Akukho okutholiwe

    I-Bioburden

    ≤50CFU/g okuqinile

    Izinsalela ze-Nucleic acid

    <5pg/mg okuqinile

     

    Izakhiwo

    Umthombo

    I-albhamu ye-Ttirachium

    Inombolo ye-EC

    3.4.21.64(I-Recombinant evela ku-albhamu ye-Ttirachium)

    Isisindo samangqamuzana

    29kDa (SDS-PAGE)

    Iphuzu le-isoelectric

    7.81 Umfanekiso

    I-pH engcono kakhulu

    7.0-12.0 (Bonke benza umsebenzi ophezulu) Fig.2

    Izinga lokushisa eliphezulu

    65℃ Fig.3

    Ukuqina kwe-pH

    pH 4.5-12.5 (25℃,16h) Fig.4

    Ukuzinza okushisayo

    Ngaphansi kuka-50℃ (pH 8.0, 30min) Fig.5

    Ukuqina kwesitoreji

    Igcinwe ku-25℃ izinyanga ezingu-12 Fig.6

    Izishoshovu

    SDS, urea

    Ama-inhibitors

    I-Diisopropyl fluorophosphate;i-benzylsulfonyl fluoride

     

    Izimo Zokugcina

    Gcina i-lyophilized powder ku -25~-15 ℃ isikhathi eside kude nokukhanya;Ngemva kokuhlakazwa, i-aliquot ibe umthamo ofanele wesitoreji sesikhathi esifushane ku-2-8℃ kude nokukhanya noma isitoreji sesikhathi eside ku--25~-15 ℃ kude nokukhanya.

     

    Izinyathelo zokuzivikela

    Gqoka amagilavu ​​okuzivikela kanye nezibuko lapho usebenzisa noma ukala, futhi uhlale unomoya omuhle ngemva kokuwusebenzisa.Lo mkhiqizo ungase ubangele ukungezwani nesikhumba kanye nokucasuka okukhulu kwamehlo.Uma uhogele, kungase kubangele ukungezwani komzimba noma izimpawu zesifuba somoya noma i-dyspnea.Ingabangela ukucasuka kokuphefumula.

     

    Incazelo yeyunithi

    Iyunithi eyodwa ye-NGS Protease K ichazwa njengenani le-enzyme edingekayo ukuze i-hydrolyze i-casein ibe yi-1 μmol L-tyrosine ngaphansi kwezimo zokunquma ezijwayelekile.

     

     Ukulungiswa kwama-reagents

    I-Reagent

    Umkhiqizi

    Ikhathalogi

    I-Casein technicalkusuka obisini lwezinkomo

    Sigma Aldrich

    C7078

    NaOH

    I-Sinopharm ChemicalInkampani Reagent Co., Ltd.

    10019762

    NaH2PO4·2H2O

    I-Sinopharm ChemicalInkampani Reagent Co., Ltd.

    20040718

    I-Na2HPO4

    I-Sinopharm ChemicalInkampani Reagent Co., Ltd.

    20040618

    I-Trichloroacetic acid

    I-Sinopharm ChemicalInkampani Reagent Co., Ltd.

    80132618

    I-sodium acetate

    I-Sinopharm ChemicalInkampani Reagent Co., Ltd.

    10018818

    I-acetic acid

    I-Sinopharm ChemicalInkampani Reagent Co., Ltd.

    10000218

    I-HCl

    I-Sinopharm ChemicalInkampani Reagent Co., Ltd.

    10011018

    I-sodium carbonate

    I-Sinopharm ChemicalInkampani Reagent Co., Ltd.

    10019260

    I-Foline-phenol

    I-Sangon Biotech (Shanghai)Co., Ltd.

    I-A500467-0100

    I-L-tyrosine

    Sigma

    93829

    I-Reagent I:

    I-Substrate: I-1% I-Casein evela kwikhambi lobisi lwezinkomo: ncibilikisa i-1g ye-bovine milk casein ku-50ml we-0.1M isisombululo se-sodium phosphate, i-pH 8.0, ukushisa emanzini okugeza ngo-65-70 °C imizuzu engu-15, gqugquzela futhi uncibilike, uphole ngamanzi, kulungiswa ngu i-sodium hydroxide iye ku-pH 8.0, bese ihlanjululwa ibe ngu-100ml.

    I-Reagent II:

    Isixazululo se-TCA: 0.1M trichloroacetic acid, 0.2M sodium acetate kanye ne-0.3M acetic acid (isisindo esingu-1.64g trichloroacetic acid + 1.64g i-sodium acetate + 1.724mL i-acetic acid ngokulandelana, engeza amanzi angama-50mL ahlanganisiwe, lungisa ne-4.HC0 ku-pH, 4. 100ml).

    I-Reagent III:

    Isixazululo se-sodium carbonate esingu-0.4m (nesisindo esingu-4.24g anhydrous sodium carbonate futhi sincibilike emanzini angu-100mL)

    I-Reagent IV:

    I-Folin phenol reagent: nciphisa izikhathi ezi-5 ngamanzi akhishwe.

    I-Reagent V:

    I-enzyme diluent: 0.1 M isisombululo se-sodium phosphate, i-pH 8.0.

    I-Reagent VI:

    Isixazululo esijwayelekile se-L-tyrosine: 0, 0.005, 0.025, 0.05, 0.075, 0.1, 0.25 umol/ml L-tyrosine ehlakazwe nge-0.2M HCl.

     

    Inqubo

    1. Vula i-spectrophotometer ye-UV-Vis bese ukhetha ukulinganisa kwe-photometric.

    2. Setha ubude begagasi njengo-660nm.

    3. Vula ukugeza kwamanzi, setha izinga lokushisa libe ngu-37℃, uqinisekise izinga lokushisa elingashintshiwe imizuzu engu-3-5.

    4. Shisisa i-substrate engu-0.5mL ku-2mL centrifuge tube endaweni yokugeza yamanzi engu-37℃ imizuzu engu-10.

    5. Khipha isixazululo se-enzyme ehlanjululwe engu-0.5mL kushubhu ye-centrifuge eshisiwe ngaphambili imizuzu eyi-10.Setha i-enzyme diluent njengeqembu elingenalutho.

    6. Engeza i-reagent ye-TCA engu-1.0 mL ngokushesha ngemva kokusabela.Hlanganisa kahle bese ufukamela emanzini okugeza imizuzu engama-30.

    7. Isixazululo se-Centrifugate reaction.

    8. Engeza izingxenye ezilandelayo ngokulandelana okushiwo.

    I-Reagent

    Ivolumu

    Onamandla

    0.5 ml

    0.4M Sodium carbonate

    2.5 mL

    I-Folin phenol reagent

    0.5 ml

    9. Hlanganisa kahle ngaphambi kokufukamela emanzini okugeza ngo-37℃ imizuzu engu-30.

    10. OD660kwanqunywa njenge-OD1;iqembu lokulawula elingenalutho: I-enzyme diluent isetshenziswa esikhundleni sesisombululo se-enzyme ukuze kunqunywe i-OD660njengoba OD2, ΔOD=OD1-OD2.

    11. Ijika elijwayelekile le-L-tyrosine: 0.5mL isixazululo se-concentration esihlukile se-L-tyrosine, 2.5mL 0.4M I-Sodium carbonate, 0.5mL i-Folin phenol reagent ku-5mL centrifuge tube, incubate ku-37℃ imizuzu engu-30, thola i-OD660ngokugxila okuhlukile kwe-L-tyrosine, kwase kutholakala ijika elijwayelekile elingu-Y=kX+b, lapho u-Y ewugxilisano we-L-tyrosine, u-X ungu-OD600.

     

    Ukubala

     

    2: Isamba sevolumu yesixazululo sokusabela (mL)

    0.5: Umthamo wesixazululo se-enzyme (mL)

    0.5: Ivolumu yoketshezi yokusabela esetshenziswa ekunqumeni kwe-chromogenic (mL)

    10: Isikhathi sokuphendula (imizuzu)

    Df: Dilution multiple

    CUkugxila kwe-enzyme (mg/mL)

    Amanani

     

    Fig.1 izinsalela ze-DNA

    Isampula

    Ave C4

    I-Nucleic Acid

    Ukubuyisela(pg/mg)

    Ukubuyisela(%)

    Ingqikithi ye-Nucleic

    I-Acid ( p/mg)

    I-PRK

    24.66

    2.23

    83%

    2.687

    I-PRK+STD2

    18.723

    126.728

    -

    -

    I-STD1

    12.955

     

     

     

     

    -

     

     

     

     

    -

     

     

     

     

    -

    I-STD2

    16

    I-STD3

    19.125

    I-STD4

    23.135

    I-STD5

    26.625

    I-RNA-Free H2O

    Akunqunyiwe

    -

    -

    -

     

    Fig.2 Optimum pH

     

    Fig.3 Izinga lokushisa eliphezulu

     

    Fig.4 pH Ukuzinza

     

    Fig.5 Ukuzinza kwe-thermal

     

    Fig.6 Ukuqina kwesitoreji ku-25℃

     

     

     

     

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