mouse monoclonal anti coup tfii  (R&D Systems)

Journal: Biomedicine & Pharmacotherapy

Article Title: Advanced cell therapy with low tissue factor loaded product NestaCell® does not confer thrombogenic risk for critically ill COVID-19 heparin-treated patients

doi: 10.1016/j.biopha.2022.112920

Figure Lengend Snippet: Results of ROTEM analysis of platelet-poor plasmas (PPPs) from four health donors (A–D) and five patients with critically ill COVID-19 (E–I) before (negative control) and after activation with ellagic acid and phospholipids (activators of intrinsic coagulation pathway - PTT) and external source of TF/CD142 (extrinsic coagulation pathway - PT), showing a significant clotting time (CT) reduction after activation, demonstrating that both intrinsic and extrinsic coagulation pathways are preserved in all analyzed PPPs. Analysis performed in triplicate.

Article Snippet: To evaluate the percentage of TF/CD142-positive, all sub-lots of hIDPSC (NestaCell® product) or HUVEC cells (used as a control for not expressing TF), at a concentration of 1.0 × 10 6 cells per 100 μL, were incubated overnight at 4°C with the mouse monoclonal anti-human TF/CD142 (Thermo Fischer Scientific, Carlsbad, USA, catalog number MA1–43029), at a dilution of 1:100.

Techniques: Negative Control, Activation Assay, Coagulation

Journal: Cancers

Article Title: Interferon-γ Produced by EBV-Positive Neoplastic NK-Cells Induces Differentiation into Macrophages and Procoagulant Activity of Monocytes, Which Leads to HLH

doi: 10.3390/cancers13205097

Figure Lengend Snippet: Procoagulant activity was enhanced by the supernatants of EBV-positive NK-cell lines. THP-1 cells and human monocytes were co-cultured with each supernatant of EBV-positive T- or NK- cell lines for 24 h. The same amount of culture media of EBV-positive T- or NK-cell lines was added as an untreated control. After the co-culture, the target cells were subjected to each assay. ( A ) The procoagulant activity (PCA) on THP-1 cells was measured by normal plasma-based recalcification time ( n = 6). The shortening of coagulation time indicates increased the PCA. ( B ) The PCA on human monocytes of three independent healthy donors was measured ( n = 3). ( C ) The mRNA expression of the tissue factor (TF) in THP-1 cells was examined by qRT-PCR ( n = 3). The expression was normalized to GAPDH mRNA. ( D ) The mRNA expression of TF in human monocytes of three independent healthy donors was measured ( n = 3). ( E ) The cell surface TF antigen on THP-1 cells was analyzed by flow cytometry using an antibody to TF or isotype-matched control. The mean fluorescent intensity of TF was normalized by that of isotype-matched control and expressed as MFIR ( n = 4). ( F ) To investigate the effects of cell surface TF on PCA, THP-1 cells were treated with mouse monoclonal anti-human TF antibody or the same amount of irrelevant IgG. After the treatment of antibody, the cell surface PCA was assessed ( n = 4). The data are shown as the mean ± SD. Statistical analyses were assessed and are indicated as * p < 0.05 and ** p < 0.01 compared to the control.

Article Snippet: After the stimulation, the target cells were analyzed by a FACSCalibur™ flow cytometer (Becton Dickinson and Company, Franklin Lakes, NJ, USA) with FITC-conjugated mouse anti-human CD11b (ab176541, abcam, Cambridge, UK), FITC-conjugated mouse anti-human CD80 (560926, BD Pharmingen, Franklin Lakes, NJ, USA), FITC-conjugated mouse anti-human CD206 (321103, Biolegend, San Diego, CA, USA), and monoclonal mouse anti-human TF IgG (ADG4509; American Diagnostics, Greenwich, CT, USA) with FITC-labeled anti-mouse IgG antibody (1030-02, SouthernBiotech, Birmingham, AL, USA).

Techniques: Activity Assay, Cell Culture, Co-Culture Assay, Coagulation, Expressing, Quantitative RT-PCR, Flow Cytometry