anti tissue inhibitor  (R&D Systems)

Journal: International Journal of Oral Science

Article Title: Transcriptome analysis of ankylosed primary molars with infraocclusion

doi: 10.1038/s41368-019-0070-1

Figure Lengend Snippet: Summary of immunostained proteins in the P1 molar tooth germ for the six genes under consideration

Article Snippet: Primary antibodies were: rabbit polyclonal anti-periostin (POSTN) (1:100, LifeSpan Biosciences, Seattle), mouse monoclonal anti-tissue inhibitor of metalloproteinases 2 (TIMP2) (1:400, Millipore, Temecula), rabbit polyclonal anti-matrix metalloproteinase 2 (MMP2) (1:50, Millipore, Temecula), mouse monoclonal anti-cytokeratin 14 (CK14) (1:200, Thermo Fisher Scientific, Rockford), mouse monoclonal anti-desmoglein 3 (DSG3) (1:100, Thermo Fisher Scientific, Rockford) and mouse monoclonal anti-plakophilin 1 (PKP1) (1:50, Abcam, Cambridge).

Techniques: Expressing

Journal: International Journal of Oral Science

Article Title: Transcriptome analysis of ankylosed primary molars with infraocclusion

doi: 10.1038/s41368-019-0070-1

Figure Lengend Snippet: Immunohistochemical staining of proteins encoded by over-expressed (CK14, DSG3, PKP1) and under-expressed (POSTN, TIMP2, MMP2) genes in infraocclusion. a Hematoxylin and Eosin staining of P1 mouse molar tooth germ in the sagittal plane with various nascent structures within the dental follicle. b CK14 was strongly stained in the dental follicle. c Higher magnification of boxed area showed CK14 immunostaining in the stellate reticulum, stratum intermedium, inner enamel epithelium and ameloblast layer. d DSG3 was strongly stained in the dental follicle and to a lesser extent in the ameloblast layer. e PKP1 was strongly stained in the dental follicle, as well as the inner enamel epithelium and the apical aspect of the ameloblast layer. f POSTN was immunostained only in the dental follicle. g MMP2 was expressed only in the dental follicle and stellate reticulum. h TIMP2 was the only protein in this sample not expressed in the dental follicle, but was abundantly present in the ameloblast and odontoblast layers. Scale bars are 100 μm

Article Snippet: Primary antibodies were: rabbit polyclonal anti-periostin (POSTN) (1:100, LifeSpan Biosciences, Seattle), mouse monoclonal anti-tissue inhibitor of metalloproteinases 2 (TIMP2) (1:400, Millipore, Temecula), rabbit polyclonal anti-matrix metalloproteinase 2 (MMP2) (1:50, Millipore, Temecula), mouse monoclonal anti-cytokeratin 14 (CK14) (1:200, Thermo Fisher Scientific, Rockford), mouse monoclonal anti-desmoglein 3 (DSG3) (1:100, Thermo Fisher Scientific, Rockford) and mouse monoclonal anti-plakophilin 1 (PKP1) (1:50, Abcam, Cambridge).

Techniques: Immunohistochemical staining, Staining, Immunostaining

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