BIOSENSORS heliX e matla hammoho le Reducing Agent Kit 1
Likarolo tsa Bohlokoa
- E lumella ho fokotsa li-bond tsa disulfide ho sebelisa TCEP.
- E lumellana le liprotheine le li-antibodies (MW> 20 kDa).
- Phokotso e khethehileng ea li-antibodies.
- Ho kopana hang-hang ha limolek'hule tse fokotsehileng hoa khoneha (sheba buka ea mosebedisi HK-MAL).
- Ho fokotsa li-ligand tse ngata ho ka etsoa ka nako e le 'ngoe.
- E kenyelletsa li-reagents bakeng sa karabelo e 'meli ea phokotso ea motho ka mong.
Tlhaloso ea Sehlahisoa
Nomoro ea odara: RK-PA-1
Tafole 1. Litaba le Boitsebiso ba Polokelo
| Lintho tse bonahalang | Cap | Chelete | Polokelo |
| TCEP (500 mM) | Khubedu | 2 x 20µL | -20°C |
| Fokotsa Buffer [1] | Bosoeu | 2 x 1.8 mL | -20°C |
| Hloekiso spin kholomo | Khubedu | 4 x | 2-8°C |
| 2.0 mL reaction tubes for purification spin column | 4 x | RT | |
| Sefate sa Centrifugal (10 kDa MWCO) | 2 x | RT | |
| Tube ea pokello ea centrifugation | 4 x | RT |
Bakeng sa tšebeliso ea lipatlisiso feela.
Sehlahisoa sena se na le nako ea nako ea shelofo, ka kopo, sheba letsatsi la ho felloa ke nako ho label.
| BOHLOKOA | Lihlahisoa li ka romelloa ka lithempereichara tse fapaneng, empa polokelo e lokela ho latela tataiso e boletsoeng ho Lethathamo. Resin slurry e kholomong ea ho hloekisa e na le 0.02% sodium azide. |
Lisebelisuoa Tse Eketsehileng Tse Hlokehang
Tafole 2. Lisebelisuoa tse Eketsehileng
| Lintho tse bonahalang | Maikutlo |
| Benchtop microcentrifuge | Lebelo le hlokahalang la lipakeng tsa 1,000 xg ho isa ho 13,000 xg |
| Pherekano | |
| 1.5 ml ea methapo ea methapo |
Litharollo tsohle tse hlokahalang le li-buffers li kenyellelitsoe ka har'a kit.
Mehato e 3 ea Phokotso ea Biomolecule ho Reaction Tube
Ka kopo bala protocol eohle pele u qala le etsa mehato yohle ntle le tshitiso.
| KELETSO | Protocol ena e ka etsoa ka nako e le 'ngoe bakeng sa liketso tse ngata tsa ho kopanya |
Buffer Exchange ea Protheine / Antibody
- Koetlisa lera la yuniti ea filthara ea centrifugal ka 100 µL Reducing Buffer.
- Kenya hoo e ka bang. 200 µg (ho fihla ho 500 µg) ea protheine/antibody ho yuniti ea sefe ho tloha mohato oa 1. Ha ho hlokahala eketsa Reducing Buffer ho sefe ho fihlela bophahamo ba bophahamo ba 450 µL bo fihletsoe le centrifuge ho 13,000 xg (ho fihla ho 14,000 xg) bakeng sa 5 min le ho lahla phallo-ho feta.
- Kenya 350 µL ea Reducing Buffer le centrifuge ho 13,000 xg bakeng sa 5 mins 'me u lahle phallo hape.
- Pheta mohato oa 3, nako e 'ngoe hape ka nako ea ho qetela ea centrifugation ea 10 min ho fumana molumo oa ho qetela oa hoo e ka bang. 80µL.
- Bokella sampe ka ba ka pipette ea 100 µL ka har'a tšupu e ncha kapa ka ho beha sesebelisoa sa filtara holimo ka har'a tube e ncha ea centrifugal, centrifuge metsotso e 2 ho 1000 x g.
Ho fokotseha ha TCEP
- Lokisetsa tharollo ea 2.5 mM TCEP ho Fokotsa Buffer ka ho eketsa 1 µL ea 500 mM TCEP ho 199 µL Reducing Buffer.
Lokisetsa kamehla ka ho toba pele o sebelisoa. - Eketsa ho fokotsa Buffer ho sample vial bakeng sa kakaretso ea 98 µL ebe u eketsa 2 µL ea 2.5 mM TCEP tharollo ho fumana motsoako oa ho qetela oa 50 µM TCEP. Kopanya karabelo ka ho phaphatha holimo le tlase.
- Eketsa tharollo ka 37 ° C ka 400 rpm bakeng sa 2 h ho netefatsa ho fokotseha ho feletseng.
Tlhoekiso le Boiketlo
- Lokisetsa litšiea tse peli tsa tlhoekiso (sekoaelo se sefubelu) bakeng sa ho hlatsoa TCEP e le 'ngoe:
a. Tlosa tiiso e ka tlase ea kholomo 'me u lokolle cap (u se ke ua tlosa cap).
b. Beha kholomo ka har'a tube ea 2.0 mL ea reaction.
c. Centrifuge ho 1,500 × g bakeng sa 1 min ho tlosa tharollo ea polokelo.
d. Kenya 400 µL ea Reducing Buffer ho bethe ea resin ea kholomo. Centrifuge ho 1,500 × g bakeng sa 1 min ho tlosa buffer.
e. Pheta mohato oa d 'me u lahle buffer e hlahisoang ho tsoa ho tšupu ea karabelo. Kholomo ea ho hloekisa e lokela ho ba boemong bo omileng.
TCEP hlatsoa
a. Beha litšiea ho tloha mohatong o fetileng ka har'a li-tubes tse ncha tsa 1.5 mL.
b. Tlosa sekepe sa spin kholomo ea nomoro ea 1 'me u tšollele s ka bonoloample holimo ho bethe ea resin.
c. Centrifuge ho 1,500 xg bakeng sa 2 min ho bokella sample (phallo-through). Lahla karolo ea ho hloekisa ka mor'a tšebeliso.
d. Tlosa kepisi ea spin kholomo ea nomoro ea 2 ebe u tšela sample ho tloha mohatong o fetileng ho ea betheng ea resin.
e. Centrifuge ho 1,500 xg bakeng sa 2 min ho bokella sample (phallo-through). Lahla karolo ea ho hloekisa ka mor'a tšebeliso.
f. Thiol coupling ho latela bukana ea mosebelisi (HK-MAL-1 kapa HK-MAL-2) ea sample ka etsoa.
Tlhahisoleseling e 'Ngoe
Buffer Exchange le Concentration le Centrifugal Filter Units
- Nka yuniti e le 'ngoe ea filthara ea centrifugal, eketsa molumo o loketseng oa buffer sesebelisoa sa sefe, ebe u se koala.
- Beha sesebelisoa sa filthara se kentsoeng ka har'a rotor ea centrifuge, u lokise leqhama la cap ho ea bohareng ba rotor; kgahlano le sesebedisoa se tshoanang le sona.
- Spina sesebelisoa ho 13,000 xg (kapa 14,000 xg) ka nako e fanoeng.
- Tlosa ho phalla 'me u phete mehato ea 1-3.
- Tlosa sesebelisoa se bokelitsoeng ho centrifuge 'me u arole sesebelisoa sa filthara ho tloha tube ea microcentrifuge.
- Ho khutlisa conjugate, beha sesebelisoa sa filthara fatše ka har'a tube e hloekileng ea centrifugal, u lokise cap e bulehileng ho ea bohareng ba rotor; kgahlano le sesebedisoa se tshoanang le sona. Spin bakeng sa metsotso ea 2 ho 1,000 xg ho fetisetsa sample ho tloha sesebelisoa ho ea ho tube.
Linomoro tsa Odara tse Molemo
Tafole 3. Linomoro tsa Order
| Lebitso la Sehlahiswa | Chelete | Odara No |
| heliX® Thiol coupling kit 1 | 5 x | HK-MAL-1 |
| heliX® Thiol coupling kit 2 | 5 x | HK-MAL-2 |
| Sefate sa Centrifugal (3 kDa MWCO) | Li-5 likhomphutha. | CF-003-5 |
| Sefate sa Centrifugal (10 kDa MWCO) | Li-5 likhomphutha. | CF-010-5 |
| 10x Buffer A [2] | 50 mL (e fana ka 500 ml) | BU-P-150-10 |
| 5x Buffer B [3] | 50 mL (e fana ka 250 ml) | BU-P-1000-5 |
Ikopanye
| Dynamic Biosensors GmbH | Litlhaloso tsa likarolo tsa Dynamic Biosensors, Inc |
| Perchtinger Str. 8/10 | 300 Trade Center, Suite 1400 |
| 81379 Munich | Woburn, MA 01801 |
| Jeremane | usa |
Lintlha tsa Odara order@dynamic-biosensors.com
Tšehetso ea tekheniki support@dynamic-biosensors.com
- Ho fokotsa Buffer: 100 mM Na2HPO4/NaH2PO4, 300 mM NaCl, 5 mM EDTA, pH 7.4
- Buffer A: 50 mM Na2HPO4/NaH2PO4, 150 mM NaCl, pH 7.2
- Buffer B: 50 mM Na2HPO4/NaH2PO4, 1 M NaCl, pH 7.2
TS'ESO MOEKETSI
www.dynamic-biosensors.com
Lisebelisoa le li-chips li entsoe le ho etsoa Jeremane.
©2024 Dynamic Biosensors GmbH | Dynamic Biosensors, Inc. Litokelo tsohle li sirelelitsoe.
Litokomane / Lisebelisoa
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BIOSENSORS heliX e matla hammoho le Reducing Agent Kit 1 [pdf] Bukana ea Mosebelisi RK-PA-1, heliX mmoho le Reducing Agent Kit 1, heliX plus, Reducing Agent Kit 1, Agent Kit 1, Kit 1 |
