I-MaxSortin® CD14 Isolation Kit

Isitoko: Sikhona
Imodeli: TL-625KIT

Xhumana nathi

Imininingwane Yomkhiqizo

Ukwakheka

Igama	Inombolo ye-athikili	Ukucaciswa	Izimo Zokugcina	I-Shelf Life
I-MaxSortin® CD14 Separation Magnetic Ubuhlalu	I-TL-625	2 mL ngesamba samaseli angu-1×10⁹	2 - 8 °C	6 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® CD14 Separation Kit ingase isetshenziselwe ukucebisa noma ukususa ama-monocyte nama-macrophage kumaseli egazi lomuntu we-peripheral mononuclear cell (PBMCs). Ngokufaka ubuhlalu obuzibuthe obuhlukanisayo be-nanoscale CD14 ngamaseli, ukuhlelwa kwamaseli e-CD14+ kungafinyelelwa. Ngokusebenzisa ubuhlalu obuzibuthe bokuhlukanisa i-MaxSortin® CD14 ukufukamela nama-PBMC bese kwenziwa ukuhlukanisa kazibuthe, amaseli e-CD14+ angahlukaniswa futhi anothiswe, adlale indima ekukhipheni noma ekuhlanzeni amaseli e-CD14+.

Imiyalo Yokusetshenziswa

Izinqubo Zokuhlola

1. Misa kabusha ama-PBMC abantu kubhafa ye-PBS equkethe u-1% we-albumin ye-human serum (HSA). Thatha isampula ukuze ubale bese udlulisela amaseli angu-1×10⁷ kushubhu ye-Ep engu-1.5 mL. Bese i-centrifuge ku-1500 rpm imizuzu emi-5.
2. Lahla amandla angaphezu kwavamile, thatha u-80 μL we-MaxSortin® Cell Sorting Buffer ukuze umise kabusha amaseli, engeza u-20 μL We-MaxSortin® CD14 Separation Magnetic Ubuhlalu, 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 engu-15 ml.
6. Ngemva kokuthi i-MaxSortin® Cell Sorting Buffer isiphumile, khipha ikholomu ehlukanisayo kusihlungi se-MACS bese usifaka kwelinye ishubhu elisha 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. 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.