Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in a mouse cell line using 4 different commercial anti-mouse TF antibodies. Proteins in cell lysates (10 μg) from the mouse pancreatic cancer cell lines KPC2 wild-type (WT) and KPC2 TF knockout (KO) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a membrane. TF was detected using 4 commercial anti-mouse TF antibodies: (A) Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody, (B) R&D Systems (R&D) AF3178 goat anti-mouse TF polyclonal antibody, (C) Abcam ab189483 rabbit anti-mouse TF monoclonal antibody, or (D) Cell Signaling Technology (Cell Signaling) 44861 rabbit anti-mouse TF monoclonal antibody. These anti-TF antibodies were detected using species-specific secondary antibodies . β-Actin was used as a loading control and detected with an anti–β-actin antibody . The binding of the anti–β-actin antibody was detected with a horse anti-mouse immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the blots were exposed for 1 second for both TF and β-actin. All blots were visualized using the iBright FL1000 imaging system. kDa, kilodalton.

Article Snippet: TF was detected using 4 commercial anti-mouse TF antibodies: (A) Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody, (B) R&D Systems (R&D) AF3178 goat anti-mouse TF polyclonal antibody, (C) Abcam ab189483 rabbit anti-mouse TF monoclonal antibody, or (D) Cell Signaling Technology (Cell Signaling) 44861 rabbit anti-mouse TF monoclonal antibody.

Techniques: Knock-Out, Polyacrylamide Gel Electrophoresis, Membrane, Control, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) using the Santa Cruz antibody. Proteins in 2 sets of lysates from mouse brain (B), heart (H), lung (Lu), and spleen (S; 40 μg) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to a membrane. The membrane was then cut into halves. One-half was incubated overnight with the Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody. The other half was incubated in 5% bovine serum albumin in Tris-buffered saline with 0.1% Tween. Both membranes were then incubated with a horse anti-mouse immunoglobulin G horseradish peroxidase-linked antibody . α-Actinin was used as a loading control and detected with an anti–α-actinin antibody . The binding of the anti–α-actinin antibody was detected with a goat anti-rabbit immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the blots were exposed for 10 seconds (TF) and 15 seconds (α-actinin). All blots were visualized using the iBright FL1000 imaging system. kDa, kilodalton.

Article Snippet: TF was detected using 4 commercial anti-mouse TF antibodies: (A) Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody, (B) R&D Systems (R&D) AF3178 goat anti-mouse TF polyclonal antibody, (C) Abcam ab189483 rabbit anti-mouse TF monoclonal antibody, or (D) Cell Signaling Technology (Cell Signaling) 44861 rabbit anti-mouse TF monoclonal antibody.

Techniques: Polyacrylamide Gel Electrophoresis, Membrane, Incubation, Saline, Control, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in mouse tissues using the R&D Systems antibody. (A) Proteins in lysates or (B) control or deglycosylated lysates from the indicated wild-type (WT) or low TF (LTF) mouse organs (40 μg) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to membranes. TF was detected using the R&D Systems AF3178 goat anti-mouse TF polyclonal antibody. The binding of the R&D Systems anti-TF antibody was detected with a rabbit anti-goat immunoglobulin G horseradish peroxidase-linked antibody . α-Actinin was used as a loading control and detected with an anti–α-actinin antibody . The binding of the anti–α-actinin antibody was detected with a goat anti-rabbit immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the TF blots were exposed for 1 second. For α-actinin, the blots were exposed for (A) 15 or (B) 30 seconds. All blots were visualized using the iBright FL1000 imaging system. ab, antibody; dgTF, deglycosylated tissue factor; gTF, glycosylated tissue factor; kDa, kilodalton.

Article Snippet: TF was detected using 4 commercial anti-mouse TF antibodies: (A) Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody, (B) R&D Systems (R&D) AF3178 goat anti-mouse TF polyclonal antibody, (C) Abcam ab189483 rabbit anti-mouse TF monoclonal antibody, or (D) Cell Signaling Technology (Cell Signaling) 44861 rabbit anti-mouse TF monoclonal antibody.

Techniques: Control, Polyacrylamide Gel Electrophoresis, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in mouse tissues using the Abcam antibody. (A) Proteins in lysates or (B) control or deglycosylated lysates from the indicated wild-type (WT) or low TF (LTF) mouse organs (40 μg) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to membranes. TF was detected using the Abcam ab189483 rabbit anti-mouse TF monoclonal antibody. The binding of the Abcam anti-TF antibody was detected with a goat anti-rabbit immunoglobulin G horseradish peroxidase-linked antibody . α-Actinin was used as a loading control and detected with an anti–α-actinin antibody . The binding of the anti–α-actinin antibody was detected with a goat anti-rabbit immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the TF blots were exposed for (A) 10, (B, top panel) 1, or (B, middle panel) 15 seconds. For α-actinin, the blots were exposed for (A) 15 or (B) 30 seconds. All blots were visualized using the iBright FL1000 imaging system. ab, antibody; dgTF, deglycosylated tissue factor; gTF, glycosylated tissue factor; kDa, kilodalton.

Article Snippet: TF was detected using 4 commercial anti-mouse TF antibodies: (A) Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody, (B) R&D Systems (R&D) AF3178 goat anti-mouse TF polyclonal antibody, (C) Abcam ab189483 rabbit anti-mouse TF monoclonal antibody, or (D) Cell Signaling Technology (Cell Signaling) 44861 rabbit anti-mouse TF monoclonal antibody.

Techniques: Control, Polyacrylamide Gel Electrophoresis, Binding Assay, Imaging

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Tissue factor detection in mouse tissues by western blotting using commercial antibodies

doi: 10.1016/j.rpth.2025.103336

Figure Lengend Snippet: Detection of tissue factor (TF) in mouse tissues using the Cell Signaling Technology antibody. (A) Proteins in lysates or (B) control or deglycosylated lysates from the indicated wild-type (WT) or low TF (LTF) mouse organs (40 μg) were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to membranes. TF was detected using the Cell Signaling Technology 44861 rabbit anti-mouse TF monoclonal antibody. The binding of the Cell Signaling Technology anti-TF antibody was detected with a goat anti-rabbit immunoglobulin G horseradish peroxidase-linked antibody . α-Actinin was used as a loading control and detected with an anti–α-actinin antibody . The binding of the anti–α-actinin antibody was detected with a goat anti-rabbit immunoglobulin G horseradish peroxidase-linked antibody . For imaging, the TF blots were exposed for (A) 2, (B, top panel) 1, or (B, middle panel) 15 seconds. For α-actinin, the blots were exposed for (A) 15 or (B) 30 seconds. All blots were visualized using the iBright FL1000 imaging system. ab, antibody; dgTF, deglycosylated tissue factor; gTF, glycosylated tissue factor; kDa, kilodalton.

Article Snippet: TF was detected using 4 commercial anti-mouse TF antibodies: (A) Santa Cruz sc-374441 mouse anti-mouse TF monoclonal antibody, (B) R&D Systems (R&D) AF3178 goat anti-mouse TF polyclonal antibody, (C) Abcam ab189483 rabbit anti-mouse TF monoclonal antibody, or (D) Cell Signaling Technology (Cell Signaling) 44861 rabbit anti-mouse TF monoclonal antibody.

Techniques: Control, Polyacrylamide Gel Electrophoresis, Binding Assay, Imaging

Journal: Molecular medicine reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation.

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: Figure 1. HDBECs (2x105) were incubated for 24 h with recombinant TF (0, 0.5 and 2 U/ml) or PAR2‑AP (SLIGKV; 20 µM), or with recombinant TF (0.5 U/ml) that was pre‑incubated for 60 min with 10H10 or HTF1 antibodies (20 µg/ml). Cells were also pre‑incubated for 60 min with AIIB2 or SAM11 antibodies (20 µg/ml), prior to addition of TF. The cells were harvested after 24 h, into separate aliquots (1x105 cells). Total RNA was isolated from one group and the mRNA quantified by RT‑qPCR, against β‑actin. Other aliquots were analysed by western blotting. The data show the amounts of (A) Inhibitor of CDK p16INKa mRNA (n=5), and (B) the relative amounts of p16INKa protein (n=3) calculated from (C) the western blots of p16INKa protein (using a goat anti‑human antibody) and against GAPDH. TF, tissue factor; PAR2, protease‑activated receptor 2.

Article Snippet: In some experiments, TF was pre‐incubated at 37 ̊C for 60 min with 10H10 antibody (20 μg/ml; cat. no. 9010‐5059; Bio‐rad laboratories, inc.) to block TF proliferative signal‐ ling via the TF exosite (50) or HTF1 antibody (20 μg/ml; cat. no. 16‐1429‐85; eBioscience; Thermo Fisher Scientific, inc.) to block the protease activity of the TF‐fViia complex (51).

Techniques: Incubation, Recombinant, Isolation, Western Blot

Journal: Molecular medicine reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation.

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: Figure 2. HDBECs (2x105) were incubated for 24 h with recombinant TF (0, 0.5 and 2 U/ml) or PAR2‑AP (SLIGKV; 20 µM), or with recombinant TF (0.5 U/ml) that was pre‑incubated for 60 min with 10H10 or HTF1 antibodies (20 µg/ml). Groups of cells were also pre‑incubated for 60 min with AIIB2 or SAM11 antibodies (20 µg/ml), prior to the addition of TF. The cells were harvested after 24 h, into separate aliquots (1x105 cells). Total RNA was isolated from one group and the mRNA quantified by RT‑qPCR, against β‑actin. Other aliquots were analysed by western blotting. The data show the amounts of (A) CDK interacting protein/Wildtype p53‑activated fragment p21CIP1/WAF1 mRNA (n=5), and (B) the relative amounts of p21CIP1/WAF1 protein (n=3) calculated from (C) the western blots of p21CIP1/WAF1 protein, (using a mouse anti‑human antibody) and against GAPDH. TF, tissue factor; PAR2, protease‑activated receptor 2.

Article Snippet: In some experiments, TF was pre‐incubated at 37 ̊C for 60 min with 10H10 antibody (20 μg/ml; cat. no. 9010‐5059; Bio‐rad laboratories, inc.) to block TF proliferative signal‐ ling via the TF exosite (50) or HTF1 antibody (20 μg/ml; cat. no. 16‐1429‐85; eBioscience; Thermo Fisher Scientific, inc.) to block the protease activity of the TF‐fViia complex (51).

Techniques: Incubation, Recombinant, Isolation, Western Blot

Journal: Molecular medicine reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation.

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: Figure 3. Examination of the influence of TF on cell cycle indicators. (A) Groups of HDBECs (1x105) were transfected with the pGL3‑promoter vector containing the E2F‑1 enhancer sequence. The cells were then treated for 24 h with recombinant TF (0, 0.5 and 2 U/ml), combinations of TF (0.5 U/ml) with the shown antibodies, or with PAR2‑AP (SLIGKV; 20 µM) and the luciferase activity was measured within 24 h (n=3). (B) Groups of cells (5x104) were seeded in 96‑well plates and treated with recombinant TF (0, 0.5 and 2 U/ml) for 24 h. The cells were then fixed and probed with an anti‑phospho‑Thr821/826 human retinoblastoma protein antibody (1:1,000 v/v) for 1 h. The samples were then incubated with an HRP‑conjugated donkey anti‑goat IgG diluted 1:3,000 v/v) for 1 h, developed with a TMB substrate and the absorptions determined at 450 nm (n=3). (C) Groups of cells (1x105) were incubated for 24 h with TF (0, 0.5 and 2 U/ml) or PAR2‑AP (20 µM), or with recombinant TF (0.5 U/ml) that was pre‑incubated for 60 min with 10H10 or HTF1 antibodies (20 µg/ml). Groups of cells were also pre‑incubated for 60 min with AIIB2 or SAM11 antibodies (20 µg/ml), prior to addition of recombinant TF. The cells were harvested after 24 h, total RNA was isolated and the amount of cyclin D1 mRNA determined against that of β‑actin (n=4). (D) Groups of cells (1x105) were treated as aforementioned and cell numbers were determined using crystal violet staining (n=3). TF, tissue factor; PAR2, protease‑activated receptor 2; E2F‑1; Early region 2 binding factor.

Article Snippet: In some experiments, TF was pre‐incubated at 37 ̊C for 60 min with 10H10 antibody (20 μg/ml; cat. no. 9010‐5059; Bio‐rad laboratories, inc.) to block TF proliferative signal‐ ling via the TF exosite (50) or HTF1 antibody (20 μg/ml; cat. no. 16‐1429‐85; eBioscience; Thermo Fisher Scientific, inc.) to block the protease activity of the TF‐fViia complex (51).

Techniques: Transfection, Plasmid Preparation, Sequencing, Recombinant, Luciferase, Activity Assay, Incubation, Isolation, Staining, Binding Assay

Journal: iScience

Article Title: Coagulation factors promote brown adipose tissue dysfunction and abnormal systemic metabolism in obesity

doi: 10.1016/j.isci.2022.104547

Figure Lengend Snippet:

Article Snippet: pCMV6-Kan/Neo-mouse F3 , OriGENE , MC200642.

Techniques: Virus, Expressing, Recombinant, Coagulation, Enzyme-linked Immunosorbent Assay, Microarray, Software