Journal: Breast cancer research : BCR

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer.

doi: 10.1186/s13058-024-01906-6

Figure Lengend Snippet: Fig. 1 Heparanase expression in immortal mammary epithelial cells and TNBC cell lines. For the western blot analysis of heparanase, (A) Adjusted volume of cell supernatants, inversely proportionate to the total protein masses in the corresponding cell lysates were probed with heparanase antibody and densitometries analyzed using the student’s t-test to determine significance. * represents p < 0.05 and ** represents p < 0.01 when comparing expression in MCF-10A cells to the expression in TNBC cell lines. (B) Equal amounts of cell lysate from each cell line were probed. Western blots were performed on two biological replicates. The densitometric analyses of the bands were calculated. The student’s t-test was performed to determine significance. * rep resents p < 0.05 and ** represents p < 0.01 when comparing expression in MCF-10A cells to the expression in TNBC cell lines. (C) Extracellular heparanase expression was determined in nontumorigenic immortal mammary epithelial MCF-10A cells and TNBC MDA-MB 231, Hs 578t, and MDA-MB 468 cells by ELISAs. The standard deviation was calculated from three independent experiments performed in triplicate. The student’s t-test was performed with * representing p < 0.05 and ** representing p < 0.01, comparing expression levels in MCF-10A to those in the TNBC cell lines. (D) Heparan sulfate expression was examined in the supernatants of TNBC cells and control immortal MCF-10A cells via ELISA analysis with MDA-MB 468 expressing the most. The stan dard deviation was calculated from three independent experiments performed in triplicate. The student’s t-test was performed to determine the signifi cance with * representing p < 0.05 and ** representing p < 0.01 comparing the protein expression in MCF-10A to the protein expressions in the TNBC cell lines. (E) and (F) ROC Plotter was applied to identify whether expression was different between chemotherapy responders (n = 30) and non-responders (n = 124). Low heparanase expression was not significantly predictive of TNBC patient chemotherapy response (ROC p = 0.06, Mann-Whitney p = 0.087)

Article Snippet: The membranes were blocked with 5% nonfat milk at room temperature for an hour and incubated overnight at 4 °C with a primary antibody: heparanase (1:200 dilution; Novus Biologicals, Cat# NBP303846, RRID: AB_2927437) diluted in 5% nonfat milk.

Techniques: Expressing, Western Blot, Standard Deviation, Control, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Breast cancer research : BCR

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer.

doi: 10.1186/s13058-024-01906-6

Figure Lengend Snippet: Fig. 2 Statistical analysis for heparanase immunohistochemistry comparing different breast cancer subtypes and normal breast tissue. For the breast cancer tissue array and slides stained for heparanase: TNBC (n = 75), ER+/PR+ (n = 18), HER2+ (n = 14), normal (n = 4), and DCIS (n = 5), (A) images were taken of heparanase-stained AMSBIO BR1202B breast cancer tissue array and normal and DCIS slides on an Evos FL Auto 2 microscope (40×). (B) A one- way ANOVA test indicated no significant difference in the H-scores of heparanase-stained cells among subtypes. A Kruskal-Wallis test indicated that there was no significant difference: (C) in the percentage of heparanase positively stained cells in the tissue sections of normal breast tissue, DCIS, and invasive breast cancer subtypes; (D) in the percentage of heparanase weakly stained cells in the tissue sections of normal breast tissue, DCIS, and invasive breast cancer subtypes; (E) in the percentage of heparanase moderately stained cells in the tissue sections of normal breast tissue, DCIS, and invasive breast cancer subtypes; (F) in the percentage of heparanase strongly stained cells in the tissue sections of normal breast tissue, DCIS, and invasive breast cancer subtypes

Article Snippet: The membranes were blocked with 5% nonfat milk at room temperature for an hour and incubated overnight at 4 °C with a primary antibody: heparanase (1:200 dilution; Novus Biologicals, Cat# NBP303846, RRID: AB_2927437) diluted in 5% nonfat milk.

Techniques: Immunohistochemistry, Staining, Microscopy

Journal: Breast cancer research : BCR

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer.

doi: 10.1186/s13058-024-01906-6

Figure Lengend Snippet: Fig. 3 Effects of heparanase inhibitor OGT 2115 and chemotherapeutic agent paclitaxel on cell viability. Percentage loss of cell viability was measured in treated (A) nontumorigenic immortal mammary epithelial MCF-10A cells and TNBC (B) MDA-MB 231, (C) Hs 578t, and (D) MDA-MB 468 cells. The treat ments applied were vehicle addition (paclitaxel, purple), heparan sodium sulfate (50 µM, teal), and OGT 2115 (20 µM, purple-red) or the combination (light blue). Heparan sodium sulfate and OGT 2115 were administered for 48 h, and paclitaxel was added for the final 6 h to replicate exposure times in patients. The standard deviation was calculated from three independent experiments performed in triplicate. One-way ANOVA with Tukey’s HSD was applied to ascertain significance. * represents p < 0.05 and ** represents p < 0.01 when comparing vehicle addition to heparan sodium sulfate, OGT 2115, or the combination

Article Snippet: The membranes were blocked with 5% nonfat milk at room temperature for an hour and incubated overnight at 4 °C with a primary antibody: heparanase (1:200 dilution; Novus Biologicals, Cat# NBP303846, RRID: AB_2927437) diluted in 5% nonfat milk.

Techniques: Standard Deviation

Journal: Breast cancer research : BCR

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer.

doi: 10.1186/s13058-024-01906-6

Figure Lengend Snippet: Fig. 4 Heparanase inhibitor OGT 2115 and chemotherapeutic agent paclitaxel influence extracellular ATP concentrations. Extracellular ATP concentra tions were measured in the supernatants of treated (A) nontumorigenic immortal mammary epithelial MCF-10 A cells and TNBC (B) MDA-MB 231, (C) Hs 578t, and (D) MDA-MB 468 cells. The treatments: vehicle addition (paclitaxel, purple), heparan sodium sulfate (50 µM, teal), and OGT 2115 (20 µM, purple-red), or the combination regimen (light blue). Heparan sodium sulfate and OGT 2115 were administered for 48 h and paclitaxel was added for the final 6 h to replicate exposure times in patients. The standard deviation was calculated from three independent experiments performed in triplicate. One- way ANOVA with Tukey’s HSD was applied to ascertain significance. * represents p < 0.05 and ** represents p < 0.01 when comparing vehicle addition to heparan sodium sulfate, OGT 2115, or the combination regimen

Article Snippet: The membranes were blocked with 5% nonfat milk at room temperature for an hour and incubated overnight at 4 °C with a primary antibody: heparanase (1:200 dilution; Novus Biologicals, Cat# NBP303846, RRID: AB_2927437) diluted in 5% nonfat milk.

Techniques: Standard Deviation

Journal: Breast cancer research : BCR

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer.

doi: 10.1186/s13058-024-01906-6

Figure Lengend Snippet: Fig. 5 Reversal of heparanase inhibitor’s effects by P2RX4 and P2RX7 inhibitors. For (A) and (B), Hs 578t cells were treated with OGT 2115 (20 µM, 48 h), paclitaxel (100 µM, the final 6 h of the 48-hour time course to replicate exposure times in patients), heparan sodium sulfate (50 µM, 48 h), A437809 (20 µM, 6 h), or a combination of the different drug agents. The standard deviation was calculated from three independent experiments performed in triplicate. One-way ANOVA with Tukey’s HSD was applied to ascertain significance. * represents p < 0.05 and ** represents p < 0.01 when comparing paclitaxel and OGT 2115 to paclitaxel, OGT 2115 and A437809 and + + represents p < 0.01 when comparing paclitaxel, OGT 2115 and heparan sulfate to paclitaxel, OGT 2115, heparan sodium sulfate and A437809. For (C) and (D) Hs 578t cells were treated with OGT 2115 (20 µM, 48 h), paclitaxel (100 µM, final 6 h of the 48-hour time course to replicate exposure times in patients), heparan sodium sulfate (50 µM, 48 h), 5-BDBD (20 µM, 6 h), or combinations. The standard deviation was calculated from three independent experiments performed in triplicate. One-way ANOVA with Tukey’s HSD was applied to ascertain sig nificance. * represents p < 0.05; ** represents p < 0.01 when comparing paclitaxel and OGT 2115 to paclitaxel, OGT 2115 and 5-BDBD and + + represents p < 0.01 when comparing paclitaxel, OGT 2115 and heparan sulfate to paclitaxel, OGT 2115, heparan sodium sulfate and 5-BDBD

Article Snippet: The membranes were blocked with 5% nonfat milk at room temperature for an hour and incubated overnight at 4 °C with a primary antibody: heparanase (1:200 dilution; Novus Biologicals, Cat# NBP303846, RRID: AB_2927437) diluted in 5% nonfat milk.

Techniques: Standard Deviation

Journal: Breast cancer research : BCR

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer.

doi: 10.1186/s13058-024-01906-6

Figure Lengend Snippet: Fig. 6 Tumorsphere formation efficiency assays for treated TNBC cells. Effects of heparanase inhibitor on cancer-initiating cells were determined through the tumorsphere formation efficiency assay in which TNBC cell lines were treated with vehicle (DMSO), paclitaxel (100 µM, final 6 h of the 48-hour time course to replicate exposure times in patients), heparan sodium sulfate (50 µM, 48 h), OGT 2115 (20 µM, 48 h), or the different combinations listed. (A) Tumorsphere images obtained (10×) are displayed for each treatment of MDA-MB 231, MDA-MB 468, and Hs 578t cells with paclitaxel, OGT 2115, heparan sodium sulfate, or the different combinations. The combination regimens showed a significant decrease in tumorsphere formation when compared to the single-agent treatments of vehicle, paclitaxel, heparan sodium sulfate, or OGT 2115 treated (B) MDA-MB 231, (C) MDA-MB 468, and (D) Hs 578t cells. Three independent experiments were performed in triplicate. One-way ANOVA with Tukey’s HSD was applied to ascertain significance. ** represents p < 0.01 when comparing paclitaxel to paclitaxel and OGT 2115. ++ represents p < 0.01 when comparing paclitaxel to paclitaxel, heparan sodium sulfate, and OGT 2115

Article Snippet: The membranes were blocked with 5% nonfat milk at room temperature for an hour and incubated overnight at 4 °C with a primary antibody: heparanase (1:200 dilution; Novus Biologicals, Cat# NBP303846, RRID: AB_2927437) diluted in 5% nonfat milk.

Techniques:

Journal: Frontiers in oncology

Article Title: miR-15b-5p Promotes Growth and Metastasis in Breast Cancer by Targeting HPSE2.

doi: 10.3389/fonc.2020.00108

Figure Lengend Snippet: FIGURE 1 | miRNA expression profiling in three paired breast cancer and adjacent normal tissues. (A) The heatmap reveals clusters of differentially expressed miRNAs; green indicates relatively low expression, and red indicates relatively high expression. (B) miRNAs–gene network was constructed to illustrate the key regulatory functions of the identified miRNAs and their target genes. The size of the circle or square node represents the degree value. A higher degree of gene/miRNAs indicates that it plays a more important role in the signaling network. (C,D) Correlation of miR-15b-5p or HPSE2 with overall survival (OS) or relapse-free survival (RFS) of breast cancer patients. The data illustrated are from the Kaplan–Meier Plotter database (http://kmplot.com/analysis/).

Article Snippet: The anti-HPSE2 antibody was purchased from Abcam Biotechnology (Cambridge, UK), and anti-GAPDH was purchased from Proteintech (Wuhan, China).

Techniques: Expressing, Construct

Journal: Frontiers in oncology

Article Title: miR-15b-5p Promotes Growth and Metastasis in Breast Cancer by Targeting HPSE2.

doi: 10.3389/fonc.2020.00108

Figure Lengend Snippet: FIGURE 2 | Expression correlation analysis of miR-15b-5p and HPSE2 in breast cancer tissues and matched adjacent non-cancerous tissues. (A) In situ hybridization (ISH) demonstrating that miR-15b-5p in breast cancer tissues was higher than that in normal tissues (n = 30, P < 0.0001). (B) HPSE2 immunoreactivity in breast cancer tissues was lower than that in normal tissues. The percentage of HPSE2 expression in breast cancer or normal tissues is shown in the figure (n = 30, P < 0.0001). The patient population represented was from Outdo Biotech Co., Ltd. (Shanghai, China). (C,D) miR-15b-5p and HPSE2 expression in normal breast and breast cancer according to the TCGA data. (E) HPSE2 proteins have lower expression in breast cancer tissues. The HPSE2 levels in six human breast cancer and paired adjacent normal tissues were measured by western blot analysis. N, paired adjacent normal tissues; T, tumor tissues. Patient information is placed in the Supplementary Materials. (****P < 0.0001).

Article Snippet: The anti-HPSE2 antibody was purchased from Abcam Biotechnology (Cambridge, UK), and anti-GAPDH was purchased from Proteintech (Wuhan, China).

Techniques: Expressing, In Situ Hybridization, Western Blot

Journal: Frontiers in oncology

Article Title: miR-15b-5p Promotes Growth and Metastasis in Breast Cancer by Targeting HPSE2.

doi: 10.3389/fonc.2020.00108

Figure Lengend Snippet: FIGURE 6 | HPSE2 is a target of miR-15b-5p. (A) The mRNA level of HPSE2 in breast cancer cells overexpressing or underexpressing miR-15b-5p. (B) The protein level of HPSE2 in breast cancer cells overexpressing or underexpressing miR-15b-5p. (C) miR-15b-5p expression is correlated with HPSE2 expression in BC from TCGA data. The r and P values are from Pearson correlation. (D) The predicted relationship between miR-15b-5p and HPSE2 via the bioinformatics prediction website. (E) The dual luciferase reporter assay demonstrated a targeted relationship between miR-15b-5p and HPSE2 in different cell lines (*P < 0.05, **P < 0.01, and ***P < 0.001).

Article Snippet: The anti-HPSE2 antibody was purchased from Abcam Biotechnology (Cambridge, UK), and anti-GAPDH was purchased from Proteintech (Wuhan, China).

Techniques: Expressing, Luciferase, Reporter Assay

Journal: Frontiers in oncology

Article Title: miR-15b-5p Promotes Growth and Metastasis in Breast Cancer by Targeting HPSE2.

doi: 10.3389/fonc.2020.00108

Figure Lengend Snippet: FIGURE 7 | miR-15b-5p/HPSE2 axis may modulate the breast cancer progression. (A) The mRNA level of HPSE2 in breast cancer cells underexpressing miR-15b-5p with or without the small interference RNA of HPSE2 (si-HPSE2). (B) The protein level of HPSE2 in breast cancer cells underexpressing miR-15b-5p with or without si-HPSE2. (C) 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays showed that the effects of miR-15b-5p downregulation on breast cancer cells in inhibiting cell proliferation were significantly reversed by co-transfection of si-HPSE2. (D) Flow cytometry showed the apoptotic rates of breast cancer cells transfected with miR-15b-5p inhibitor and/or si-HPSE2. (E) The invasion of breast cancer cells underexpressing miR-15b-5p with or without si-HPSE2 (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001).

Article Snippet: The anti-HPSE2 antibody was purchased from Abcam Biotechnology (Cambridge, UK), and anti-GAPDH was purchased from Proteintech (Wuhan, China).

Techniques: Cotransfection, Flow Cytometry, Transfection

Journal: Archives of biochemistry and biophysics

Article Title: Effect of CCS on the accumulation of FALS SOD1 mutant-containing aggregates and on mitochondrial translocation of SOD1 mutants: implication of a free radical hypothesis.

doi: 10.1016/j.abb.2011.02.014

Figure Lengend Snippet: Fig. 1. Effect of simultaneous expression of FALS hSOD1 and hCCS on SOD1- containing HMWS in AAV293 cells. (A) Schematic representation of plasmid vectors used to produce dicistronic mRNA for hSOD1 and hCCS co-expression. B, BamHI; X, XhoI; N, NcoI; K, KpnI; RI, EcoRI. (B) Detection of mutant SOD1 with or without CCS revealed with reducing gel (see Materials and methods). Top panel shows a hSOD1 blot visualized by ECL chemiluminescence. Asterisks depict the high molecular- weight species. The CCS proteins were also detected by the sheep polyclonal hSOD1 (574597) antibody due to the high homology of the two proteins. Middle panel shows the identical SOD1 blot as the top panel, except with shorter exposure time. Lower panel represents a CCS blot. This figure shows typical results obtained with three independent experiments.

Article Snippet: The primary antibodies used were as follows: sheep polyclonal anti-human Cu,Zn SOD1 (574597) (1:2000; Calbiochem, La Jolla, CA, USA), rabbit polyclonal anti-human Cu,Zn SOD1 (RDI-SODabRx) (1:5000; Fitzgerald, Concord, MA, USA), mouse monoclonal anti-mitochondrial Hsp70 (MN3-028) (1:5000; Affinity Bioreagents, Golden, CO), mouse monoclonal anti-human CCS (2A1) (1:5000; Abcam, Cambridge, MA, USA), mouse monoclonal anti-actin (1:10,000; Sigma), and mouse monoclonal anti-Hsp70 (W27) (1:1000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Expressing, Plasmid Preparation, Mutagenesis, High Molecular Weight

Journal: Archives of biochemistry and biophysics

Article Title: Effect of CCS on the accumulation of FALS SOD1 mutant-containing aggregates and on mitochondrial translocation of SOD1 mutants: implication of a free radical hypothesis.

doi: 10.1016/j.abb.2011.02.014

Figure Lengend Snippet: Fig. 2. Co-expression of CCS reduced the non-ionic detergent-insoluble fraction of FALS A4V and G85R SOD1 mutants. As described in Materials and methods, ‘‘PBS- soluble fraction’’ (lane 1), ‘‘non-ionic detergent-soluble fraction’’ (lane 2), and ‘‘non- ionic detergent-insoluble fractions’’ (lane 3) were obtained and analyzed. Proteins (5 lg) were loaded onto 10–20% gradient gels. Blots were obtained with rabbit polyclonal antibody (RDI-SODabRx) to Cu,Zn SOD1, or with monoclonal antibody to CCS, and visualized by ECL chemiluminesence.

Article Snippet: The primary antibodies used were as follows: sheep polyclonal anti-human Cu,Zn SOD1 (574597) (1:2000; Calbiochem, La Jolla, CA, USA), rabbit polyclonal anti-human Cu,Zn SOD1 (RDI-SODabRx) (1:5000; Fitzgerald, Concord, MA, USA), mouse monoclonal anti-mitochondrial Hsp70 (MN3-028) (1:5000; Affinity Bioreagents, Golden, CO), mouse monoclonal anti-human CCS (2A1) (1:5000; Abcam, Cambridge, MA, USA), mouse monoclonal anti-actin (1:10,000; Sigma), and mouse monoclonal anti-Hsp70 (W27) (1:1000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Expressing

Journal: Archives of biochemistry and biophysics

Article Title: Effect of CCS on the accumulation of FALS SOD1 mutant-containing aggregates and on mitochondrial translocation of SOD1 mutants: implication of a free radical hypothesis.

doi: 10.1016/j.abb.2011.02.014

Figure Lengend Snippet: Fig. 5. Complexes formed between G85R and CCS, and between SOD1 and Hsp70 detected by co-immunoprecipitation. AAV293 cells were transiently transfected with (A and C) G85R-GFP or G85R-CCS vectors and with (B) FLAG-tagged WT SOD1 or A4V mutant expression vectors with or without CCS co-expression. At 48 h post- transfection, cells were lysed in lysis buffer (20 mM Hepes, 1% Triton X-100, 10% glycerol, pH 7.4) that contained protease inhibitor cocktail (Sigma). After brief sonication, cell lysates were centrifuged (10,000g for 5 min) and the supernatant was collected. Rabbit polyclonal antibody that does not cross-react with CCS, against SOD1 (RDI-SODabRx) (A) or mouse monoclonal antibody against Hsp70 (MN3-028) (B and C) were added to a supernatant aliquot containing 500 lg of cellular proteins and incubated for 1 h at 4 C. To isolate the SOD1 antibody conjugated proteins (A) or the Hsp70 antibody conjugated proteins (B and C), protein A-conjugated agarose beads (Pierce) were added and the mixture incubated for another 1 h at 4 C. After washing with wash buffer, the immunoprecipitates were recovered with 1 SDS sample buffer, separated by SDS–PAGE, and probed with rabbit anti-SOD1 (RDI-SODabRx), mouse monoclonal anti-CCS (2AI) and anti- Hsp70 antibodies (MN3-028) as indicated. IB, immunoblotting; IP, immunoprecipitation.

Article Snippet: The primary antibodies used were as follows: sheep polyclonal anti-human Cu,Zn SOD1 (574597) (1:2000; Calbiochem, La Jolla, CA, USA), rabbit polyclonal anti-human Cu,Zn SOD1 (RDI-SODabRx) (1:5000; Fitzgerald, Concord, MA, USA), mouse monoclonal anti-mitochondrial Hsp70 (MN3-028) (1:5000; Affinity Bioreagents, Golden, CO), mouse monoclonal anti-human CCS (2A1) (1:5000; Abcam, Cambridge, MA, USA), mouse monoclonal anti-actin (1:10,000; Sigma), and mouse monoclonal anti-Hsp70 (W27) (1:1000; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Immunoprecipitation, Transfection, Mutagenesis, Expressing, Lysis, Protease Inhibitor, Sonication, Incubation, SDS Page, Western Blot

Journal: Carbohydrate polymers

Article Title: Polymeric Fluorescent Heparin as One-Step FRET Substrate of Human Heparanase

doi: 10.1016/j.carbpol.2018.10.071

Figure Lengend Snippet: Generic structure of heparin (R = H or SO3−; R1 = H, COCH3 or SO3−) showing typical site (↓) cleaved by human heparanase (HPSE).

Article Snippet: Antibodies used for analysis include anti-heparanase 1 (aa 301-331, Boster bio, PB9427) and anti-HPSE (aa34-115, LS Bio, LS-C294467).

Techniques:

Journal: Carbohydrate polymers

Article Title: Polymeric Fluorescent Heparin as One-Step FRET Substrate of Human Heparanase

doi: 10.1016/j.carbpol.2018.10.071

Figure Lengend Snippet: FRET-based assay for human heparanase. Heparin-DE (1 mg/ml) was incubated with HPSE (1 μM) at 37°C in 20 mM sodium acetate buffer, pH 5.0. Fluorescence emission spectra (λEX = 340 nm) at (bold line) 0 h, (dotted line) 4 h.

Article Snippet: Antibodies used for analysis include anti-heparanase 1 (aa 301-331, Boster bio, PB9427) and anti-HPSE (aa34-115, LS Bio, LS-C294467).

Techniques: Incubation, Fluorescence

Journal: Carbohydrate polymers

Article Title: Polymeric Fluorescent Heparin as One-Step FRET Substrate of Human Heparanase

doi: 10.1016/j.carbpol.2018.10.071

Figure Lengend Snippet: Optimization of heparanase assay using heparin-DE as substrate. (A) Labeled heparin (1 mg/ml) was incubated 1 μM HPSE at 37 °C for indicated times followed by measurement of emission at 500 nm. (B) Labeled heparin (1 mg/ml) was incubated with varying concentrations HPSE at 37 °C for 4 h followed by fluorescence measurement. (C) Michaelis–Menten kinetics of HPSE cleavage of heparin–DE using the optimized FRET quenching assay. The cleavage reactions were performed in microplate format (100 μL) in 20 mM sodium acetate buffer, pH 5.0, containing 1 mg/mL heparin–DE and 1 μM HPSE at 37 °C for 4 h. Solid lines represent curve fitting to the standard Michaelis equation. Error bars show variation from at least 3 measurements. F and F0 are fluorescence signals corresponding to the test sample and blank, respectively.

Article Snippet: Antibodies used for analysis include anti-heparanase 1 (aa 301-331, Boster bio, PB9427) and anti-HPSE (aa34-115, LS Bio, LS-C294467).

Techniques: Labeling, Incubation, Fluorescence

Journal: Carbohydrate polymers

Article Title: Polymeric Fluorescent Heparin as One-Step FRET Substrate of Human Heparanase

doi: 10.1016/j.carbpol.2018.10.071

Figure Lengend Snippet: Suramin inhibition of HPSE using the one-step FRET quenching assay. The experiments were performed in microplate format (100 μL) in 20 mM sodium acetate buffer, pH 5.0, containing 1 mg/mL heparin–DE and 1 μM HPSE at 37 °C for 4 h in the presence of varying concentrations of suramin. Solid lines represent curve fitting to standard dose-response equation.

Article Snippet: Antibodies used for analysis include anti-heparanase 1 (aa 301-331, Boster bio, PB9427) and anti-HPSE (aa34-115, LS Bio, LS-C294467).

Techniques: Inhibition

Journal: Carbohydrate polymers

Article Title: Polymeric Fluorescent Heparin as One-Step FRET Substrate of Human Heparanase

doi: 10.1016/j.carbpol.2018.10.071

Figure Lengend Snippet: Expression of active heparanase by MCF7 cells under normoxic conditions. Inset shows the HEK cells expressed HPSE activity measurement. Results are presented as the mean±SD (n>3).

Article Snippet: Antibodies used for analysis include anti-heparanase 1 (aa 301-331, Boster bio, PB9427) and anti-HPSE (aa34-115, LS Bio, LS-C294467).

Techniques: Expressing, Activity Assay