[R&D Systems] FlowX Human NK Cell Killing Flow Cytometry Kit | 고마바이오텍

특징 

• 타겟 세포에 작용하는 Natural Killer Cell의 세포 사멸 능력을 유세포 분석 방법으로 확인하기 위한 kit입니다.
• Natural Killer (NK) cells과 Target cells을 표지하기 위한 염료, cell viability를 확인하기 위한 염료, 정량화를 위한 cell count particles, 3가지 형광처리 항체, NK 세포가 표적세포를 사멸시키는 것을 모니터링 하기 위한 유세포 분석용 염색 및 fixation buffer가 제공됩니다. 
• 세포 독성을 평가하기 위한 LAMP-1 항체가 함께 제공됩니다. 
• CAR-NK 혹은 NK Cells에 적용할 수 있습니다.
• 방사성 동위원소를 사용하지 않아 사용이 수월합니다. 

Graphical representation of killing by NK cell effectors. 

Gating for the Mito Mark dye histograms is done as follows: gate on All cells, then gate on the non-NKs (Janelia Fluor 646 negative/CD56 Alex Fluor 700 negative). This ensures that NKs are not included in the Mito Mark histograms. The graph depicts K562 killing normalized to K562 cells alone (0:1). The relative percentage of NK killing is shown at 0.5:1 and 5:1 Effector to Target ratios (see histograms).

CAR-NKs as Effective Killers.

Human NK cells were enriched from PBMCs and transduced with CD19 CAR (CAR-NK). CAR-NK expression was determined using recombinant human CD19-Atto 647N (ATM9269) and hCD56 PE Monoclonal Antibody (R&D Systems®, # FAB24086P). CAR-NKs were labeled with Janelia Fluor, Daudi target cells or K562 target cells were

labeled with Mito Mark Green, and cells were co-incubated for 2 hours at Effector to Target ratios. Anti-LAMP PE Antibody (R&D Systems, # IC4800P) was included in the culture during the 2 hour killing assay. Cells were harvested and stained with hCD56 and hCD3.