mouse anti-human vegf monoclonal antibody  (Servicebio Inc)

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Expression levels of hyaluronidase (Hyal)-1 (A) , CD44 (B) and receptor for hyaluronan-mediated motility (RHAMM) (C) in the retinal lysates of non-diabetic control rats (C) (n=12) and diabetic rats (D) (n=12) were determined by Western blot analysis. After determination of the intensity of the protein bands, intensities were adjusted to those of β-actin in the samples. Oxidative stress was monitored with the use of 2’,7’-Dichlorofluorescein (DCF) fluorescence intensity analysis (D) . Results are expressed as mean ± standard deviation. Ultra-Low molecular weight hyaluronan (ULMW-HA) induces breakdown of blood-retinal barrier (E) . ULMW-HA was injected intravitreally at the dose of 50 ng in 5 µL in one eye and the same volume of phosphate-buffered saline (PBS) was injected in the contralateral eye of normal rats. The BRB was quantified with the fluorescein isothiocyanate-conjugated dextran technique. Results are expressed as mean ± standard deviation of 12 rats. *p < 0.05 compared to the values obtained from PBS-injected eyes. (independent t-test). Western blot analysis of retinas demonstrated that intravitreal injection of ULMW-HA induced significant upregulation of the expression of phospho-NF-κB (F) , phospho-ERK1/2 (G) , vascular endothelial growth factor (VEGF) (H) , intercellular adhesion molecule-1 (ICAM-1) (I) , vascular cell adhesion molecule-1 (VCAM-1) (J) and high-mobility group box-1 (HMGB1) (K) . Results are expressed as mean ± standard deviation or standard error of mean of 8–10 rats in each group (*p < 0.05; independent t-test).

Article Snippet: To determine the presence of Hyal-1, Hyal-2, HAS2, CD44, syndecan-1, heparan sulphate and RHAMM in the vitreous samples, equal volumes (10 μL) of vitreous samples were boiled in Laemmli’s sample buffer (1:1, v/v) under reducing condition for 10 min. Immunodetection was performed with the use of rabbit polyclonal anti-Hyal-1 antibody (1:1000, NBP2-16906, Novus Biologicals), mouse polyclonal anti-Hyal-2 antibody (1:1000, H00008692-B02P, Novus Biologicals), mouse monoclonal anti-HAS2 antibody (1:1000, ab140671, Abcam), rabbit monoclonal anti-CD44 antibody (1:1000, ab189524, Abcam), rabbit monoclonal anti-RHAMM antibody (1:1000, ab124729, Abcam), rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1000, MAB1018, R&D Systems), rabbit polyclonal anti-p65 subunit of nuclear factor-kappa B (phospho-NF-κB) (1:1000, NB100-82086, Novus Biologicals), rabbit polyclonal anti-high-mobility group box1 (HMGB1) (1:1000, Cat. no. ab18256, Abcam), mouse monoclonal anti-VEGF antibody (1:750, MAB293, R&D Systems), mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:100, sc-8439, Santa Cruz Biotechnology Inc.), and mouse monoclonal anti-vascular cell adhesion molecule-1 (VCAM-1) antibody (1:100, sc-13160, Santa Cruz Biotechnology Inc.).

Techniques: Expressing, Control, Western Blot, Fluorescence, Standard Deviation, Molecular Weight, Injection, Saline

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal Müller glial cells were left untreated or treated with ultra-low molecular weight hyaluronan (ULMW-HA) (50 µg/mL) for 24 (h) (A) Protein expression of phospho-ERK1/2 and phospho-NFκB in cell lysates was determined by Western blot analysis. Levels of high mobility group box-1 (HMGB1) were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate (*p < 0.05; independent t-test). (B) Human retinal Müller glial cells were left untreated or treated with ULMW-HA, ULMW-HA plus BAY11-7085 (5 µM) or (C) ULMW-HA plus U-0126 (5 µM). Levels of vascular endothelial growth factor (VEGF), angiopoietin and monocyte chemotactic protein-1 (MCP-1/CCL2) were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation or standard error of mean from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three groups and two groups, respectively. *p < 0.05 compared with values obtained from untreated cells; #p < 0.05 compared with ULMW-HA plus BAY11–7085 or U-0126 treated cells. (D, E) Human retinal Müller glial cells were left untreated or treated with high glucose (HG) (25 mM), cobalt chloride (CoCl 2 ) (300 µM) or tumor necrosis factor-α (TNF-α) (5 ng/mL) with or without apigenin (10 µg/mL) for 24 (h) For HG treatment, cultures containing 25 mM mannitol were used as a control. Levels of monocyte chemotactic protein-1 (MCP-1/CCL2) (D) and vascular endothelial growth factor (VEGF) (E) were quantified in the culture media by ELISA. The results are expressed as mean ± standard deviation from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells. #p < 0.05 compared with values obtained from stimulated cells.

Article Snippet: To determine the presence of Hyal-1, Hyal-2, HAS2, CD44, syndecan-1, heparan sulphate and RHAMM in the vitreous samples, equal volumes (10 μL) of vitreous samples were boiled in Laemmli’s sample buffer (1:1, v/v) under reducing condition for 10 min. Immunodetection was performed with the use of rabbit polyclonal anti-Hyal-1 antibody (1:1000, NBP2-16906, Novus Biologicals), mouse polyclonal anti-Hyal-2 antibody (1:1000, H00008692-B02P, Novus Biologicals), mouse monoclonal anti-HAS2 antibody (1:1000, ab140671, Abcam), rabbit monoclonal anti-CD44 antibody (1:1000, ab189524, Abcam), rabbit monoclonal anti-RHAMM antibody (1:1000, ab124729, Abcam), rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1000, MAB1018, R&D Systems), rabbit polyclonal anti-p65 subunit of nuclear factor-kappa B (phospho-NF-κB) (1:1000, NB100-82086, Novus Biologicals), rabbit polyclonal anti-high-mobility group box1 (HMGB1) (1:1000, Cat. no. ab18256, Abcam), mouse monoclonal anti-VEGF antibody (1:750, MAB293, R&D Systems), mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:100, sc-8439, Santa Cruz Biotechnology Inc.), and mouse monoclonal anti-vascular cell adhesion molecule-1 (VCAM-1) antibody (1:100, sc-13160, Santa Cruz Biotechnology Inc.).

Techniques: Molecular Weight, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal microvascular endothelial cells (HRMECs) were left untreated or treated with high glucose (HG) (25 mM) (A) , cobalt chloride (CoCl 2 ) (300 µM) (B) or tumor necrosis factor-α (TNF-α) (5 ng/mL) (C) with or without apigenin (10 µg/mL). For HG treatment, cultures treated with mannitol (25 mM) were used as a control. Levels of soluble syndecan-1 were quantified in the culture media by ELISA. Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells. #p < 0.05 compared with values obtained from cells treated with HG, CoCl 2 or TNF-α. HRMECs were left untreated or were stimulated with ultra-low molecular weight – hyaluronan (ULMW-HA) (50 µg/mL) for 24 (h). Protein expression of phospho-ERK1/2 in the cell lysates was determined by Western blot analysis (D) . Results are expressed as mean ± standard deviation from three different experiments each performed in triplicate (*p < 0.05; independent t-test). A scratch was performed in confluent monolayers of overnight starved HRMECs with a micropipette tip subsequently, the cultures were left untreated or treated either with VEGF (10 ng/mL) or with ULMW-HA (100 µg/mL) for 16 (h) Cells were visualized using an inverted microscope. Two independent experiments were performed. Each experiment was done in duplicate, and 2–3 independent field images were taken for the migration analysis which was done by using Image J software. In the Figure, one representative image is illustrated, and the bar graphs show the analysis of all the images from each group represented as fold-change in migration versus control (E) . Results are expressed as mean ± standard deviation. One-way ANOVA and independent t-test were used for comparisons between three and two groups, respectively. *p < 0.05 compared with values obtained from control cells.

Article Snippet: To determine the presence of Hyal-1, Hyal-2, HAS2, CD44, syndecan-1, heparan sulphate and RHAMM in the vitreous samples, equal volumes (10 μL) of vitreous samples were boiled in Laemmli’s sample buffer (1:1, v/v) under reducing condition for 10 min. Immunodetection was performed with the use of rabbit polyclonal anti-Hyal-1 antibody (1:1000, NBP2-16906, Novus Biologicals), mouse polyclonal anti-Hyal-2 antibody (1:1000, H00008692-B02P, Novus Biologicals), mouse monoclonal anti-HAS2 antibody (1:1000, ab140671, Abcam), rabbit monoclonal anti-CD44 antibody (1:1000, ab189524, Abcam), rabbit monoclonal anti-RHAMM antibody (1:1000, ab124729, Abcam), rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1000, MAB1018, R&D Systems), rabbit polyclonal anti-p65 subunit of nuclear factor-kappa B (phospho-NF-κB) (1:1000, NB100-82086, Novus Biologicals), rabbit polyclonal anti-high-mobility group box1 (HMGB1) (1:1000, Cat. no. ab18256, Abcam), mouse monoclonal anti-VEGF antibody (1:750, MAB293, R&D Systems), mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:100, sc-8439, Santa Cruz Biotechnology Inc.), and mouse monoclonal anti-vascular cell adhesion molecule-1 (VCAM-1) antibody (1:100, sc-13160, Santa Cruz Biotechnology Inc.).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Standard Deviation, Molecular Weight, Expressing, Western Blot, Inverted Microscopy, Migration, Software

Journal: Frontiers in Cell and Developmental Biology

Article Title: Syndecan-1 as a predictor of vulnerable atherosclerotic plaques

doi: 10.3389/fcell.2024.1415788

Figure Lengend Snippet: Analysis of destabilizing components in atherosclerotic plaque. (A) The expression of lipid deposition in aortic root plaques was detected by oil red O with a scale bar of 100 μm, immunofluorescence detection of CD68 expression (macrophage marker) and VEGF in aortic root plaques with a scale bar of 20 μm, and IHC detection of MMP-9 expression in aortic root plaques with a scale bar of 100 μm. (B–E) Statistical chart for quantitative analysis of oil red o staining, CD68, VEGF and MMP-9 in the aortic root plaques (n = 6, one-way ANOVA). (F) Immunofluorescence detection of BODIPY and VEGF expression in LCCA plaques with a scale bar of 100 μm. IHC detection of CD68 and MMP-9 expression in LLCCA plaques with a scale bar of 100 μm. (G–J) Statistical chart for quantitative analysis of BODIPY staining, VEGF, CD68 and MMP-9 in the LCCA plaques (n = 6, unpaired t -test). * P < 0.05, ** P < 0.01, *** P < 0.001, compared with CD group; # P < 0.05, ## P < 0.01, ### P < 0.001, compared with CD + PLCA group; && P < 0.01, &&& P < 0.001, compared to HFD group.

Article Snippet: After blocking, sections were incubated with a rabbit anti-human CD68 (1:100, bs-4819R, BIOSS, China), α-SMA (1:100, A1011, Abclonal, China) and mouse anti-human VEGF (1:100, TA500289, Origene, United States of America) antibodies overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Marker, Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Syndecan-1 as a predictor of vulnerable atherosclerotic plaques

doi: 10.3389/fcell.2024.1415788

Figure Lengend Snippet: Summary of the plaque components in HFD and PLCA apoE −/− mice.

Article Snippet: After blocking, sections were incubated with a rabbit anti-human CD68 (1:100, bs-4819R, BIOSS, China), α-SMA (1:100, A1011, Abclonal, China) and mouse anti-human VEGF (1:100, TA500289, Origene, United States of America) antibodies overnight at 4°C.

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Syndecan-1 as a predictor of vulnerable atherosclerotic plaques

doi: 10.3389/fcell.2024.1415788

Figure Lengend Snippet: Serum SDC1 is an independent predictor of high-risk vulnerable plaque. (A) Serum SDC1; (B) S1P; (C) VEGF-A was detected using ELISA after 16 weeks of modeling, n = 6, one-way ANOVA. (D) ROC curve. (E) Multifactor binary logistic regression analysis was performed on serum SDC1, S1P and VEGF-A. (F) Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic accuracy of the variables. The Jorden index, calculated as sensitivity minus (1 - specificity), was used to determine the optimal cut-off point on the ROC curve. The area under the ROC curve (AUC) was utilized to assess the clinical prediction value of SDC1. Notes: AUC, area under the curve; PV+, positive predictive value; PV-, negative predictive value. * P < 0.05, * P < 0.01, *** P < 0.001, compared with CD group; # P < 0.05, compared with CD + PLCA group; && P < 0.01, compared to HFD group.

Article Snippet: After blocking, sections were incubated with a rabbit anti-human CD68 (1:100, bs-4819R, BIOSS, China), α-SMA (1:100, A1011, Abclonal, China) and mouse anti-human VEGF (1:100, TA500289, Origene, United States of America) antibodies overnight at 4°C.

Techniques: Enzyme-linked Immunosorbent Assay, Diagnostic Assay

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: The effects of anti-lung cancer in nude mice by a fully human single-chain antibody against associated antigen Ts7TMR between A549 cells and Trichinella spiralis .

doi: 10.1080/21691401.2024.2347377

Figure Lengend Snippet: Figure 2. The effect of anti-Ts7TMR scFv on A549 lung cancer and its preliminary mechanism. A. The effect of anti-Ts7TMR scFv on the proliferation of A549 cells in vitro. A549 cells were co-incubated with different doses of anti-Ts7TMR scFv (5, 25, and 45 μg) for 48h to detect cell proliferation by CCK-8 assay. B. The effect of anti-Ts7TMR scFv on A549 lung cancer growth in nude mice. Nude mice were injected subcutaneously with A549 cells and then treated subcutaneously with PBS, negative serum (Ns), anti-T. spiralis serum (Ts), and different doses of anti-Ts7TMR scFv (25, 50, and 100 μg) around the tumour for 10 consecutive days. C. Scoring of PCNA, BCL-2, and VEGF expression in tumour tissues by immunohistochemistry (400 ×). Expression of PCNA, BCL-2, and VEGF in tumours treated with PBS, negative serum (Ns), anti-T. spiralis serum (Ts), and different doses of anti-Ts7TMR scFv (25, 50, and 100 μg). Scale bar = 20 μm. *p < 0.05, **p < 0.01.

Article Snippet: Then mouse anti-human PCNA monoclonal antibody (60097-1-Ig, Proteintech), mouse anti-human BCL-2 monoclonal antibody (60178-1-Ig, Proteintech), and mouse anti-human VEGF monoclonal antibody (66828-1-Ig, Proteintech) were added separately dropwise and incubated at 37 °C for 30 min. Then the sections were incubated with biotin-conjugated Goat anti-mouse IgG at 37 °C for 30 min. Next, Streptavidin Immunohistochemistry kits (KIT-9710, maxim biotechnologies) and diaminobenzidine (dAB-0031, maxim biotechnologies) were added dropwise in sequence.

Techniques: In Vitro, Incubation, CCK-8 Assay, Injection, Expressing, Immunohistochemistry