human vegf elisa kit 271 picokine  (Boster Bio)

Journal: Frontiers in Immunology

Article Title: Physicochemical and biological characterization of a bispecific antibody in a CrossMab/KIH format that targets EGFR and VEGF-A

doi: 10.3389/fimmu.2025.1659966

Figure Lengend Snippet: Physiochemical characterization of anti-EGFR/VEGF-A CrossMab/KIH BsAb. (A) The schematic representation of anti-EGFR/VEGF-A BsAb with CrossMab/KIH format illustrates how the BsAb was constructed from four antibody fragments derived from faricimab and cetuximab and was designed to target EGFR and VEGF-A. (B) Structural integrity was evaluated via SDS-PAGE analysis of 2 µg anti-EGFR/VEGF-A BsAb, 2 µg faricimab (FAR), and 2 µg cetuximab (CET) protein samples under non-reducing and reducing conditions. (C) Representative chromatograms of anti-EGFR/VEGF-A BsAb (15 µg) and faricimab (15 µg) protein samples were generated using SEC-HPLC method (D) Representative electropherograms of anti-EGFR/VEGF-A BsAb and faricimab protein samples were generated using non-reducing and reducing CE-SDS method. Data presented in this figure represented at least three independent experiments.

Article Snippet: Human VEGF ELISA Kit Picokine ® (Cat# 0593) was purchased from Boster Bio (Pleasanton, CA, USA) and was used to quantitate the VEGF secreted from human OC cells in cell culture media.

Techniques: Construct, Derivative Assay, SDS Page, Generated

Journal: Frontiers in Immunology

Article Title: Physicochemical and biological characterization of a bispecific antibody in a CrossMab/KIH format that targets EGFR and VEGF-A

doi: 10.3389/fimmu.2025.1659966

Figure Lengend Snippet: Potency characterization of anti-EGFR/VEGF-A BsAb. (A) Dose-dependent binding activity of anti-EGFR/VEGF-A BsAb, bevacizumab, faricimab, and cetuximab was evaluated using an ELISA binding assay. (B) Simultaneous binding activity of anti-EGFR/VEGF-A BsAb to its targets, biotinylated EGFR and VEGF-A was evaluated using an ELISA binding assay. (C) Dose-dependent inhibition of VEGF-A/VEGFR2 activation by anti-EGFR/VEGF-A BsAb, bevacizumab, faricimab, and ramucirumab was performed using a VEGF activity bioassay.

Article Snippet: Human VEGF ELISA Kit Picokine ® (Cat# 0593) was purchased from Boster Bio (Pleasanton, CA, USA) and was used to quantitate the VEGF secreted from human OC cells in cell culture media.

Techniques: Binding Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Inhibition, Activation Assay, Bioassay

Journal: Frontiers in Immunology

Article Title: Physicochemical and biological characterization of a bispecific antibody in a CrossMab/KIH format that targets EGFR and VEGF-A

doi: 10.3389/fimmu.2025.1659966

Figure Lengend Snippet: Thermal stability evaluation of anti-EGFR/VEGF-A BsAb. Anti-EGFR/VEGF-A BsAb (15 µg) was stressed at 42°C for 2 weeks. The protein samples were collected at seven different time points: 0 h (Day 0), 24 h (Day 1), 48 h (Day 2), 72 h (Day 3), 168 h (Day 7), 240 h (Day 10), and 336 h (Day 14). (A) Overlaid chromatograms of thermal stressed and unstressed anti-EGFR/VEGF-A BsAb protein samples was generated using a SEC-HPLC method. (B) SDS-PAGE analysis showed the structural integrity of thermal stressed and unstressed anti-EGFR/VEGF-A BsAb protein samples under non-reducing and reducing conditions. (C) Dose-dependent binding activity of anti-EGFR/VEGF-A BsAb protein samples to EGFR and VEGF-A was evaluated using the ELISA binding assay. (D) Dose-dependent inhibition of VEGF-A/VEGFR2 activation by thermal stressed and unstressed anti-EGFR/VEGF-A BsAb protein samples was performed using a VEGF activity bioassay.

Article Snippet: Human VEGF ELISA Kit Picokine ® (Cat# 0593) was purchased from Boster Bio (Pleasanton, CA, USA) and was used to quantitate the VEGF secreted from human OC cells in cell culture media.

Techniques: Generated, SDS Page, Binding Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Inhibition, Activation Assay, Bioassay

Journal: Frontiers in Immunology

Article Title: Physicochemical and biological characterization of a bispecific antibody in a CrossMab/KIH format that targets EGFR and VEGF-A

doi: 10.3389/fimmu.2025.1659966

Figure Lengend Snippet: Inhibition of ligand-induced activation of EGFR by anti-EGFR/VEGF-A BsAb. (A) Western blot analysis was performed to measure EGFR expression levels in HUVEC and OC cell lines: PA-1, CaOV3, OVCAR3, and SKOV3 cells. Whole cell lysates were prepared from each cell line, and Western blotting was performed to measure relative EGFR protein levels in these cell lines. (B) Phospho-EGFR and EGFR levels were measured using Western blot analysis of serum-starved WCL collected from CaOV3 and SKOV3 cells. Cells were pre-treated with 10 µg indicated monoclonal antibodies and BsAb for 1 hour followed by 100 ng/mL ligand stimulation for 15 minutes.

Article Snippet: Human VEGF ELISA Kit Picokine ® (Cat# 0593) was purchased from Boster Bio (Pleasanton, CA, USA) and was used to quantitate the VEGF secreted from human OC cells in cell culture media.

Techniques: Inhibition, Activation Assay, Western Blot, Expressing, Bioprocessing

Journal: Frontiers in Immunology

Article Title: Physicochemical and biological characterization of a bispecific antibody in a CrossMab/KIH format that targets EGFR and VEGF-A

doi: 10.3389/fimmu.2025.1659966

Figure Lengend Snippet: Inhibition of ligand-induced activation of VEGFR2 by anti-EGFR/VEGF-A BsAb. (A) Western blot analysis was performed to measure VEGFR2 expression levels in HUVEC and OC cell lines: PA-1, CaOV3, OVCAR3, and SKOV3 cells. WCL were prepared from each cell line, and Western blot was performed to measure relative VEGFR2 protein levels in these cell lines. (B) Phospho-VEGFR2 and VEGFR2 levels were measured using western blot analysis of serum-starved WCL collected from CaOV3 and PA-1 cells. Cells were pre-treated with 10 µg indicated monoclonal antibodies and BsAb for 1 hour followed by 100 ng/mL ligand stimulation for 15 minutes.

Article Snippet: Human VEGF ELISA Kit Picokine ® (Cat# 0593) was purchased from Boster Bio (Pleasanton, CA, USA) and was used to quantitate the VEGF secreted from human OC cells in cell culture media.

Techniques: Inhibition, Activation Assay, Western Blot, Expressing, Bioprocessing

Journal: Frontiers in Immunology

Article Title: Physicochemical and biological characterization of a bispecific antibody in a CrossMab/KIH format that targets EGFR and VEGF-A

doi: 10.3389/fimmu.2025.1659966

Figure Lengend Snippet: Inhibition of paracrine VEGFR2 activation in HUVECs by anti-EGFR/VEGF-A BsAb. (A) The CellTiter-Glo luminescent cell viability assays were performed in HUVEC cells. 10,000 cells were seeded in white bottom of 96-well plates and allowed to adhere overnight in media with 1% FBS. After treatments with 10 μg/mL cetuximab (B) , bevacizumab (C) , or anti-EGFR/VEGF-A BsAb (D) , CellTiter-Glo reagent was added into the plates and luminescence (i.e., viability) was measured using a Promega GloMax Discover plate reader. (E) Western blot analysis was performed to measure the inhibition of ligand-induced activation of VEGFR2 and its downstream pathways (Akt, and FAK) by bevacizumab (BEV) and anti-EGFR/VEGF-A BsAb. (F) The levels of VEGF-A were determined by ELISA assay in supernatants of CaOV3, SKOV3, OVCAR3, and PA-1 cells after serum-starving the cells in a 6-well-plate for 48 h (G) Inhibition of the conditional media (CM)-mediated VEGFR2 activity in HUVEC cells by cetuximab (CET), bevacizumab (BEV), and anti-EGFR/VEGF-A BsAb. The conditional media (CM) samples were collected from SKOV3 cell culture after 48 h serum-starvation. HUVEC cells were serum-starved for 24 h CM samples were collected from the SKOV3 cells and pre-incubated with 10 μg/mL cetuximab (CET), bevacizumab (BEV), or anti-EGFR/VEGF-A BsAb. HUVEC media was removed from the cells and replaced with the CM for 2 h before WCL was harvested. Whole cell lysates were then subjected to western blot analysis.

Article Snippet: Human VEGF ELISA Kit Picokine ® (Cat# 0593) was purchased from Boster Bio (Pleasanton, CA, USA) and was used to quantitate the VEGF secreted from human OC cells in cell culture media.

Techniques: Inhibition, Activation Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Activity Assay, Cell Culture, Incubation

Journal: Frontiers in Neuroscience

Article Title: Gut microbiota-derived acetate promotes long-term recovery through angiogenesis guided by lymphatic ingrowth in older adults with stroke

doi: 10.3389/fnins.2024.1398913

Figure Lengend Snippet: (A) Heatmap of Spearman rank correlation analysis between gut microbiota and clinical indexes in the Old and Ctrl groups at the genus level. Red indicates a positive correlation, and blue indicates a negative correlation. * P < 0.05, ** P < 0.01, *** P < 0.001 for comparisons between the Old and Ctrl groups. (B) Graph comparing individual SCFA levels between the Old and Ctrl groups. P -values are as follows: P = 0.0246, P = 0.3254, P = 0.2444, P = 0.7661, P = 0.1540, P = 0.1777, P = 0.0557, and P = 0.1344 for acetate through heptanoic acid, respectively. (C) Comparison of VEGF and VEGF-C levels in serum between the two groups. (D) Spearman correlation analysis among VEGF, VEGF-C, and SCFAs. Red indicates a positive correlation, and blue indicates a negative correlation. The value in each cell represents the correlation coefficient (r). Acetate, propionate, isobutyrate, and butyrate showed varying degrees of positive correlation with VEGF and VEGF-C, while the other SCFAs did not.

Article Snippet: The concentrations of VEGF and VEGFC in serum were measured using commercially available ELISA kits: Human VEGF Quick ELISA Kit (96T, Boster Biotech Co., Ltd., China, FEK0539), Human VEGF-C ELISA Kit (96T, Boster Biotech Co., Ltd., China, EK0588), and Mouse VEGF ELISA Kit (96T, Boster Biotech Co., Ltd., China, FEK0541).

Techniques: Comparison

Journal: Frontiers in Neuroscience

Article Title: Gut microbiota-derived acetate promotes long-term recovery through angiogenesis guided by lymphatic ingrowth in older adults with stroke

doi: 10.3389/fnins.2024.1398913

Figure Lengend Snippet: (A) In the schematic brain diagram, the orange area represents the infarct area. Red boxes indicate the regions where immunofluorescence images were taken. The left side shows the ingrowth of lymphatic vessels on day 28, past stroke. Scale bar: 50 μm. (B) The distribution of meningeal lymphatic vessels on day 3 post-stroke in young FTG mice. The white arrow indicates the growth of lymphatic vessels. R: right side, L: left side. Scale bar: 1,000 μm. (C) Representative Western blot images of VEGF/GAPDH and Ang1/GAPDH in the cortex on day 28 post-stroke. (D) Quantification of VEGF/GAPDH and Ang1/GAPDH relative expression on day 28 post-stroke ( n = 5 per group). (E) Representative Western blot images of PROX1/GAPDH, Lyve-1/GAPDH, and VEGFC/β-actin in the cortex on day 28 post-stroke ( n = 5 per group). (F) Quantification of PROX1/GAPDH, Lyve-1/GAPDH, and VEGFC/β-actin relative expression on day 28 post-stroke.* P < 0.05, ** indicates a p- value of <0.01 in the aged-FTG vs. young-FTG group.

Article Snippet: The concentrations of VEGF and VEGFC in serum were measured using commercially available ELISA kits: Human VEGF Quick ELISA Kit (96T, Boster Biotech Co., Ltd., China, FEK0539), Human VEGF-C ELISA Kit (96T, Boster Biotech Co., Ltd., China, EK0588), and Mouse VEGF ELISA Kit (96T, Boster Biotech Co., Ltd., China, FEK0541).

Techniques: Immunofluorescence, Western Blot, Expressing