Human Coagulation Factor III/Tissue Factor Antibody

Detection of Human Coagulation Factor III by Western Blot. Western blot shows lysates of THP-1 human acute monocytic leukemia cell line treated (+) with TNF-a. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Coagulation Factor III/Tissue Factor Monoclonal Antibody (Catalog # MAB2339) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Coagulation Factor III at approximately 50 and 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Coagulation Factor III/Tissue Factor by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line and A549 human lung carcinoma cell line (negative control). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Coagulation Factor III/Tissue Factor Monoclonal Antibody (Catalog # MAB2339) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Coagulation Factor III/Tissue Factor at approximately 48 kDa (as indicated). GAPDH (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of Coagulation Factor III/Tissue Factor by Western Blot SC1 inhibits TF-dependent tumor cell migration(A-C) MDA-MB-231 and BxPC3 cells transfected with TRIPZ non-targeting ShRNA (Sh-NT) or TF ShRNA (Sh-TF) were induced with doxycycline (Dox) for 5-7 days to deplete TF. The cells were subjected to immunoblotting (A) or cell migration assay. Migrated cells were stained with crystal violet (B) and the quantified results are plotted (C). (D-F) The indicated cell lines were assayed similarly for cell migration with various doses of SC1 or IgG (D, E) or 100 nM SC1 (F). Quantified results are plotted. *, P<0.05; **, P<0.01; ***, P<0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28938620), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Coagulation Factor III/Tissue Factor by Western Blot Mechanistic target of rapamycin (mTOR) inhibitor differentially targets TF-mRNA transcription or TF protein degradation in a cellular context-dependent manner. (A,B) U87MG and HCC827 (A), H1975 and PC9 (B) cells were treated as indicated, subjected to qPCR analysis of TF mRNA as described in section “Materials and Methods.” The relative mRNA levels are shown. (C,D) H1975 (C) and PC9 (D) cells were treated with 5 μmol/L MTI-31 alone or in combination with 10 μmol/L CQ or 0.01 μmol/L PS-341 for 24 h, then immunoblotted (left panels) and quantified (right panels). *P < 0.05; **P < 0.01, ***P < 0.001. (E) The amino acid sequence of TF was download from https://www.ncbi.nlm.nih.gov/to set up the model of TF structure using Pymol software. The KFERQ-motif is marked in yellow and its sequence is marked in red. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Coagulation Factor III/Tissue Factor by Western Blot Mechanistic target of rapamycin (mTOR) promotes TF overexpression in EGFR-mut tumors. (A) mTOR inhibitor WYE-132 and AZD8055 decreased TF expression in four indicated cell lines. The indicated cells were treated for 24 h in full growth medium followed by immunoblotting. (B) The indicated cells were serum-starved then stimulated with 50 ng/mL EGF without or with inhibitor for 24 h. (C) Serum-starved U87MG cells were stimulated with 10% serum without or with WYE-132 for 12 h and immunoblotted. (D,E) The indicated cell lines were infected with pGIPZ-shRNA encoding lentivirus, incubated for 6–7 days in full-growth medium and immunoblotted. (F) Cells as in D and E were serum-starved, stimulated with EGF and immunoblotted. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Coagulation Factor III/Tissue Factor by Western Blot Mechanistic target of rapamycin (mTOR) inhibitor differentially targets TF-mRNA transcription or TF protein degradation in a cellular context-dependent manner. (A,B) U87MG and HCC827 (A), H1975 and PC9 (B) cells were treated as indicated, subjected to qPCR analysis of TF mRNA as described in section “Materials and Methods.” The relative mRNA levels are shown. (C,D) H1975 (C) and PC9 (D) cells were treated with 5 μmol/L MTI-31 alone or in combination with 10 μmol/L CQ or 0.01 μmol/L PS-341 for 24 h, then immunoblotted (left panels) and quantified (right panels). *P < 0.05; **P < 0.01, ***P < 0.001. (E) The amino acid sequence of TF was download from https://www.ncbi.nlm.nih.gov/to set up the model of TF structure using Pymol software. The KFERQ-motif is marked in yellow and its sequence is marked in red. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Coagulation Factor III/Tissue Factor by Western Blot Mechanistic target of rapamycin (mTOR) promotes TF overexpression in EGFR-mut tumors. (A) mTOR inhibitor WYE-132 and AZD8055 decreased TF expression in four indicated cell lines. The indicated cells were treated for 24 h in full growth medium followed by immunoblotting. (B) The indicated cells were serum-starved then stimulated with 50 ng/mL EGF without or with inhibitor for 24 h. (C) Serum-starved U87MG cells were stimulated with 10% serum without or with WYE-132 for 12 h and immunoblotted. (D,E) The indicated cell lines were infected with pGIPZ-shRNA encoding lentivirus, incubated for 6–7 days in full-growth medium and immunoblotted. (F) Cells as in D and E were serum-starved, stimulated with EGF and immunoblotted. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Coagulation Factor III/Tissue Factor by Western Blot Mechanistic target of rapamycin (mTOR) promotes TF overexpression in EGFR-mut tumors. (A) mTOR inhibitor WYE-132 and AZD8055 decreased TF expression in four indicated cell lines. The indicated cells were treated for 24 h in full growth medium followed by immunoblotting. (B) The indicated cells were serum-starved then stimulated with 50 ng/mL EGF without or with inhibitor for 24 h. (C) Serum-starved U87MG cells were stimulated with 10% serum without or with WYE-132 for 12 h and immunoblotted. (D,E) The indicated cell lines were infected with pGIPZ-shRNA encoding lentivirus, incubated for 6–7 days in full-growth medium and immunoblotted. (F) Cells as in D and E were serum-starved, stimulated with EGF and immunoblotted. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.