CytoSelect™ Leukocyte-Endothelium Adhesion Assay Kit
Leukocyte extravasation은 inflammatory diseases에 중요한 역할을 하며, 크게 Rolling, Firm adhesion, Transmigration과 같이 3가지 단계로 나눌 수 있습니다.
이러한 과정은 endothelial cell adhesion molecules과 leukocytes 특정 ligand 간의 상호작용에 의해 연속적으로 일어나게 되며, CytoSelect™ Leukocyte-endothelium Adhesion Assay는 leukocyte-endothelium interactions을 quantitative하게 측정할 수 있는 제품입니다.
- Leukocyte-endothelium interactions을 quantitative하게 측정할 수 있습니다.
- CytoTracker Solution과 TNF α만 -20℃에 보관하고 나머지 구성품은 4℃에 보관합니다.
- 100 assays에 사용할 충분한 양의 시약이 제공됩니다.
- 제품 구성
- 500 X LeukoTracker™ Solution (Part No.12101): One Tube - 100 μL
- Gelatin Solution(Part No.12102): One Bottle - 12.0 mL of sterile 0.1 % Gelatin in 1X PBS
- 4 X Lysis Buffer(Part No.12103): One Bottle - 10.0 mL
- 10 X Wash Buffer(Part No.12104): One Bottle - 20.0 mL
- TNFα (Part No. 12105): One Tube - 100 μL of 10 μg/mL TNFα in sterile 1X PBS/0.1%BSA
Figure 1. Quantitation of Human Monocytic THP-1. LeukoTracker™ labeled THP-1 cells were titrated in 1X PBS, then subsequently lysed with 2X Lysis Buffer (75 μL of cell suspension was mixed with 75 μL of 2X Lysis Buffer). Fluorescence was quantified as described in Assay Protocol.
Figure 2. Human Monocytic THP-1 Adhesion to HUVEC Monolayer. HUVEC monolayer in 48-well plate was treated with 1 μM PMA for 12 hrs. LeukoTracker™ labeled THP-1 cells were allowed to attach to HUVEC monolayer for 1 hr. Adherent cells were lysed and quantified by as described in the Assay Protocol.
Figure 3. Cytokine Activation of Human Monocytic THP-1 Adhesion to HUVEC. HUVEC monolayer (left) or THP-1 cells (right) were treated with 50 ng/mL TNFα or 1 μM PMA for 12 hrs.
LeukoTracker™ labeled THP-1 cells (50,000 cells/well) were allowed to attach to HUVEC monolayer for 1 hr. Adherent cells were lysed and quantified by as described in the Assay Protocol.