I-MaxSortin® CD3 Isolation Kit

Isitoko: Sikhona
Imodeli: TL-622KIT

Xhumana nathi

Imininingwane Yomkhiqizo

Ukwakheka

Igama	Inombolo ye-athikili	Ukucaciswa	Izimo Zokugcina	I-Shelf Life
I-MaxSortin® CD3 Separation Magnetic Ubuhlalu	I-TL-622	2 mL ngesamba samaseli angu-1×10⁹	2 - 8 °C	7 izinyanga
I-MaxSortin® Cell Sorting Buffer	I-MS-BF100	100 ml	2 - 8 °C	Izinyanga ezingu-12
Ikholomu Yokuhlukanisa ye-MaxSortin® L	I-MS-CL01	1 isiqephu	10 - 35 °C	Izinyanga ezingu-12

Imininingwane Yomkhiqizo

Izinhlobo Ezisebenzayo: Umuntu

I-Magnetic Bead Endotoxin:

Ukubukeka kwe-Magnetic Bead: Uketshezi olunsundu

Ukusetshenziswa Okuhlosiwe

I-MaxSortin® CD3 Separation Kit ingasetshenziswa ukuhlunga amaseli e-CD3+ T omuntu. Ngokufaka ubuhlalu obuzibuthe obuhlukanisa i-nanoscale CD3 ngamaseli, ukuhlungwa kwamaseli e-CD3+ kungafinyelelwa.

Imiyalo Yokusetshenziswa

Izinqubo Zokuhlola

1. Phinda umise amaseli egazi lomuntu we-peripheral mononuclear (PBMCs) kubhafa ye-PBS equkethe u-1% we-albumin ye-human serum (HSA). Thatha isampula yokubala bese udlulisela amaseli angu-1×10⁷ kushubhu ye-EP engu-1.5 mL. Bese i-centrifuge ku-1500 rpm imizuzu emi-5.
2. Thatha u-80 μL we-MaxSortin® Cell Sorting Buffer ukuze umise kabusha amaseli, engeza u-20 μL wobuhlalu obuzibuthe bokuhlukaniswa kwe-CD3, uhlanganise kahle bese ububeka esiqandisini ngo-2 - 8 °C ukuze kufakwe incubation imizuzu engu-15.
3. Thatha Ikholomu Yokuhlukanisa I-MaxSortin® L bese uyibeka kusihlungi seseli esisebenza zibuthe (i-MACS), bese uyihlanza kabili nge-1 mL ye-MaxSortin® Cell Sorting Buffer.
4. Thatha isampula eqede ukufukamela uyikhiphe esiqandisini ngo-2 - 8 °C, engeza u-1 mL we-MaxSortin® Cell Sorting Buffer, i-centrifuge ku-1500 rpm imizuzu emi-5, bese ulahla amandla amakhulu.
5. Engeza u-1 mL we-MaxSortin® Cell Sorting Buffer ukuze umise kabusha amaseli, engeza isampula kukholamu yokuhlukanisa. Ngemva kokuba igeleze ngokwemvelo, engeza i-MaxSortin® Cell Sorting Buffer ngezingxenye ezimbili, 1 mL isikhathi ngasinye. Qoqa ukungcola ngeshubhu le-centrifuge elingu-15 mL.
6.Ngemva kokuthi i-MaxSortin® Cell Sorting Buffer isiphumile, susa ikholomu yokuhlukanisa kusihlungi se-MACS bese usibeka kwelinye elisha ishubhu le-centrifuge elingu-15 mL. Engeza u-3 mL we-MaxSortin® Cell Sorting Buffer kukholamu yokuhlukanisa futhi usebenzise ipiston eza nekholomu yokuhlukanisa ukuze uphushe uketshezi luphume ngokuqondile.
7. Beka ishubhu le-centrifuge elingu-15 mL eliqukethe uketshezi oluqoqiwe ku-centrifuge evundlile kanye ne-centrifuge ku-1500 rpm imizuzu emi-5.
8. Ngemva kwe-centrifugation, thela i-supernatant, thatha i-1 mL ye-1× ye-Dulbecco's phosphate-buffered saline (DPBS) isisombululo sokumisa kabusha amaseli, ubale amaseli futhi uqhube ukutholwa kwe-cytometry yokugeleza.

Izinyathelo zokuzivikela

1. Uma ufukamela ubuhlalu kazibuthe ngamaseli, kuyadingeka ukuxutshwa kahle ukuze kuthuthukiswe ukusebenza kahle kokuhlunga.

2. Uma usebenzisa lo mkhiqizo ukususa nokuhlunga amaseli e-CD3+, kuyadingeka ukwandisa ngokufanelekile inani lobuhlalu obukazibuthe obusetshenzisiwe.

3. Ikholomu yebhafa neyokwehlukanisa efakwe kule khithi ingasetshenziselwa isilingo sokuqala. Uma imisebenzi yokuhlola eyengeziwe ihilelekile, sicela uthenge i-MaxSortin® Cell Sorting Buffer (Inombolo Yesihloko: MS-BF100) kanye Nekholomu Yokuhlukaniswa Kwe-MaxSortin® (Inombolo Yesihloko: MS-CL01) ngokuhlukana.