human mouse tissue inhibitor  (Cusabio)

Journal: Journal of Hematology & Oncology

Article Title: TFPIα and TFPIβ are expressed at the surface of breast cancer cells and inhibit TF-FVIIa activity

doi: 10.1186/1756-8722-6-5

Figure Lengend Snippet: Characterization of TFPI and TF in a selection of tumor derived breast cancer cell lines and normal cells

Article Snippet: The polyclonal rabbit anti-human tissue factor pathway inhibitor antibody (4901/ADG72), monoclonal mouse anti-human tissue factor pathway inhibitor antibody (4903), and monoclonal mouse anti-human tissue factor antibody (ADG4508) were from American Diagnostica (Greenwich, CT, USA), while the rabbit-IgG-UNLB and goat anti-rabbit IgG (H+L)-RPE antibodies were from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: Selection, Derivative Assay, In Vitro, Transformation Assay

Journal: Journal of Hematology & Oncology

Article Title: TFPIα and TFPIβ are expressed at the surface of breast cancer cells and inhibit TF-FVIIa activity

doi: 10.1186/1756-8722-6-5

Figure Lengend Snippet: TFPI expression in different breast cancer cell lines and normal cells. Cells were seeded in 6-wells trays to reach ~80% confluence in three days before media were collected and cells lysed for RNA isolation. ( A ) Relative TFPIα and TFPIβ mRNA expression measured using qRT-PCR. ΔΔCt values were calculated using 18s rRNA as an endogenous control and the TFPI negative, non-human CHO cells as a negative control. ( B ) Secreted TFPIα protein measured in cell media and normalized to total protein amounts. Mean values + SD (n = 3) are presented.

Article Snippet: The polyclonal rabbit anti-human tissue factor pathway inhibitor antibody (4901/ADG72), monoclonal mouse anti-human tissue factor pathway inhibitor antibody (4903), and monoclonal mouse anti-human tissue factor antibody (ADG4508) were from American Diagnostica (Greenwich, CT, USA), while the rabbit-IgG-UNLB and goat anti-rabbit IgG (H+L)-RPE antibodies were from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: Expressing, Isolation, Quantitative RT-PCR, Control, Negative Control

Journal: Journal of Hematology & Oncology

Article Title: TFPIα and TFPIβ are expressed at the surface of breast cancer cells and inhibit TF-FVIIa activity

doi: 10.1186/1756-8722-6-5

Figure Lengend Snippet: PI-PLC treatment of Sum102 and MDA-MB-231 cells. Serum-starved cells were treated with SFM (untreated) or PI-PLC (1 U/mL) for 2 hours before the supernatant was removed and cells lysed or collected for flow cytometry analysis. Total and free TFPI antigen measured in the supernatant ( A and F ) and lysate ( B ) of untreated and PI-PLC treated Sum102 ( A and B ) and MDA-MB-231 ( F ) cells. Mean values + SD (n ≥ 8) of three to six independent experiments are presented. All statistical comparisons were conducted between PI-PLC treated samples and untreated controls. Flow cytometry analysis of untreated (black, solid) and PI-PLC treated (black, broken) Sum102 ( C ) and MDA-MB-231 ( G ) cells. One representative experiment of three is shown. ( D ) Fluorescence images of SFM + glycerol (untreated, left panel) and PI-PLC (middle panel) treated Sum102 cells stained with antibody against both isoforms of TFPI. The right panel shows a secondary antibody background control staining. One representative experiment of three is shown. ( E ) Western blot of supernatants from SFM + glycerol (untreated) and PI-PLC treated Sum102 cells. Samples were deglycosylated and incubated with a polyclonal anti-human TFPI antibody.

Article Snippet: The polyclonal rabbit anti-human tissue factor pathway inhibitor antibody (4901/ADG72), monoclonal mouse anti-human tissue factor pathway inhibitor antibody (4903), and monoclonal mouse anti-human tissue factor antibody (ADG4508) were from American Diagnostica (Greenwich, CT, USA), while the rabbit-IgG-UNLB and goat anti-rabbit IgG (H+L)-RPE antibodies were from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: Flow Cytometry, Fluorescence, Staining, Control, Western Blot, Incubation

Journal: Journal of Hematology & Oncology

Article Title: TFPIα and TFPIβ are expressed at the surface of breast cancer cells and inhibit TF-FVIIa activity

doi: 10.1186/1756-8722-6-5

Figure Lengend Snippet: Heparin treatment of Sum102 and MDA-MB-231 cells. Serum-starved cells were treated with SFM (untreated) or heparin (5 U/mL) for 2 hours or with PI-PLC + heparin (2+2 hours) before the supernatant was removed and cells lysed or collected for flow cytometry analysis. The supernatant was discarded after PI-PLC pre-treatment. Free and total TFPI antigen measured in the supernatant ( A and C ) and lysate ( B and D ) of untreated, heparin or PI-PLC + heparin treated Sum102 ( A and B ) and MDA-MB-231 ( C and D ) cells. Mean values + SD (n ≥ 9) of three to six independent experiments are presented. Statistical comparisons were conducted between heparin/PI-PLC+heparin treated samples and untreated controls. ( E ) Flow cytometry analysis of untreated (black, solid) and heparin treated (black, dotted) Sum102 (left panel) and MDA-MB-231 (right panel) cells. One representative experiment of three is shown.

Article Snippet: The polyclonal rabbit anti-human tissue factor pathway inhibitor antibody (4901/ADG72), monoclonal mouse anti-human tissue factor pathway inhibitor antibody (4903), and monoclonal mouse anti-human tissue factor antibody (ADG4508) were from American Diagnostica (Greenwich, CT, USA), while the rabbit-IgG-UNLB and goat anti-rabbit IgG (H+L)-RPE antibodies were from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: Flow Cytometry

Journal: Journal of Hematology & Oncology

Article Title: TFPIα and TFPIβ are expressed at the surface of breast cancer cells and inhibit TF-FVIIa activity

doi: 10.1186/1756-8722-6-5

Figure Lengend Snippet: PI-PLC and heparin treatment of HCAEC cells. Serum-starved cells were treated with SFM (untreated), PI-PLC (1 U/mL), or heparin (5U/mL) for 2 hours or with PI-PLC + heparin (2+2 hours) before the supernatant was removed and cells lysed or collected for flow cytometry analysis. The supernatant was discarded after PI-PLC pre-treatment. Total and free TFPI antigen measured in the supernatant ( A and C ) and cell lysate ( B and D ) of untreated/PI-PLC ( A and B ) and untreated/heparin/PI-PLC+heparin ( C and D ) treated HCAEC cells. Mean values (n ≥ 5; supernatant, n = 4; cell lysate) + SD of two independent experiments. Statistical comparisons were conducted between PI-PLC or heparin/PI-PLC+heparin treated samples and untreated controls. ( E ) Flow cytometry analysis of untreated (black, solid), PI-PLC (black, broken, left panel), and heparin (black, dotted, right panel) treated HCAEC cells. One representative experiment of three is shown.

Article Snippet: The polyclonal rabbit anti-human tissue factor pathway inhibitor antibody (4901/ADG72), monoclonal mouse anti-human tissue factor pathway inhibitor antibody (4903), and monoclonal mouse anti-human tissue factor antibody (ADG4508) were from American Diagnostica (Greenwich, CT, USA), while the rabbit-IgG-UNLB and goat anti-rabbit IgG (H+L)-RPE antibodies were from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: Flow Cytometry

Journal: Journal of Hematology & Oncology

Article Title: TFPIα and TFPIβ are expressed at the surface of breast cancer cells and inhibit TF-FVIIa activity

doi: 10.1186/1756-8722-6-5

Figure Lengend Snippet: Cell surface anticoagulant activity of PI-PLC treated cells. Confluent Sum102, MDA-MB-231, and HCAEC cells grown in 24-wells trays were treated with SFM (untreated), PI-PLC, monoclonal anti-TFPI antibody, or anti-TF antibody for 2 hours before the supernatant was removed and cells were washed and assayed for TF activity. Sum102 ( A ), MDA-MB-231 ( B ), and HCAEC ( C ) cells were incubated with FVIIa and FX for 1 hour before the reaction was stopped and a substrate to FXa was added. The results were analyzed colorimetrically at 405 nm. The amount of FXa generated is displayed as mean mU/mL + SD (n ≥ 7) of three to four independent experiments. Statistical comparisons were conducted between PI-PLC treated samples and untreated controls. ( D ) Fluorescence images of SFM + Glycerol (untreated, upper panels) and PI-PLC (bottom panels) treated Sum102 cells stained with antibodies against both isoforms of TFPI (green, left panels) and TF (red, middle-left panels). After merging the images, co-localization of the two antibodies was indicated by a yellow color (middle-right panels). Nuclear staining with DAPI is depicted in the left panels. One representative experiment of three is shown.

Article Snippet: The polyclonal rabbit anti-human tissue factor pathway inhibitor antibody (4901/ADG72), monoclonal mouse anti-human tissue factor pathway inhibitor antibody (4903), and monoclonal mouse anti-human tissue factor antibody (ADG4508) were from American Diagnostica (Greenwich, CT, USA), while the rabbit-IgG-UNLB and goat anti-rabbit IgG (H+L)-RPE antibodies were from Southern Biotechnology Associates (Birmingham, AL, USA).

Techniques: Activity Assay, Incubation, Generated, Fluorescence, Staining