mouse anti human vegf c 1  (Santa Cruz Biotechnology)

Journal: Circulation Research

Article Title: Overexpression of VEGF 165 b, an Inhibitory Splice Variant of Vascular Endothelial Growth Factor, Leads to Insufficient Angiogenesis in Patients With Systemic Sclerosis

doi: 10.1161/circresaha.111.242057

Figure Lengend Snippet: Figure 2. Overexpression of VEGF165b in SSc skin determined by immunofluorescence and confocal microscopy. A, Blots of SDS–polyacryl- amide gels loaded with recombinant human (rh) VEGF165b and rh VEGF165 were probed with anti-VEGF165b and anti-pan-VEGF antibodies. Anti- VEGF165b antibody specifically recognizes VEGF165b only. Anti-pan-VEGF antibodies detect both VEGF165b and VEGF165. B–H, Representative micro- photographs of VEGF165b immunostaining in skin sections. In control skin, VEGF165b was constitutively expressed in the epidermis (e) and dermis (B, C). In the majority of control subjects, VEGF165b expres- sion was weak (B). Keratinocytes, fibroblasts (arrow- head), and capillary vessels (arrow) of 6 of 23 con- trol subjects showed an intense VEGF165b immunopositivity (C). In SSc skin, a strong expres- sion of VEGF165b was evident in the epidermis (e), fibroblasts (arrowheads), microvascular endothelial cells (arrows), and perivascular inflammatory cells (*) (D-F). G, In SSc skin, endothelial cells and vascular smooth muscle cells of an arteriole (arrow) strongly expressed VEGF165b. H, Intense immunopositivity for VEGF165b in a large inflammatory infiltrate of SSc hypodermis (*). D, G, and H show early SSc skin, E and F show late SSc skin. I and J, Double immuno- staining for VEGF165b (green) and CD31 (red) in SSc skin. Original magnification, 63. K, Densitometric analysis of VEGF165b immunofluorescent staining. Data are meanSD of optical density in arbitrary units (a.u.). *P0.001 vs control.

Article Snippet: This antibody has been shown using Western blotting and immunohistochemistry to specifically detect the VEGF165b isoform of VEGF, but not the pro-angiogenic VEGF165 isoform, even when present at 10,000-fold greater concentrations.9,12 Protein samples extracted from various human tissues, when probed with this antibody, revealed immunoblotting bands of the same size as that detected using a pan-VEGF antibody.9 Three commercially available anti-VEGF-A antibodies were used to investigate the expression of all isoforms of VEGF (including VEGF165b) – termed pan-VEGF: rabbit anti-human VEGF antibody A-20 and mouse anti-human VEGF antibody C-1 (both from Santa Cruz Biotechnology, Santa Cruz, CA, USA; cat. no. sc-152 and sc-7269, respectively), and mouse anti- human VEGF antibody (BD PharMingen, Heidelberg, Germany; cat. no. 555036).

Techniques: Over Expression, Confocal Microscopy, Recombinant, Immunostaining, Control, Staining

Journal: Circulation Research

Article Title: Overexpression of VEGF 165 b, an Inhibitory Splice Variant of Vascular Endothelial Growth Factor, Leads to Insufficient Angiogenesis in Patients With Systemic Sclerosis

doi: 10.1161/circresaha.111.242057

Figure Lengend Snippet: Figure 3. VEGF165b and pan- VEGF expression in skin biopsy samples. A, Serial skin sections from SSc patients immunostained with anti- VEGF165b–specific antibodies (upper panels) and anti-pan- VEGF antibodies (lower pan- els). In epidermis (e) and der- mis, VEGF165b-specific and pan-VEGF stains showed simi- lar intensity in the same tissue structures. Asterisks indicate a dermal perivascular inflamma- tory infiltrate. B, Double immu- nostaining for VEGF165b (green) and pan-VEGF (red) in SSc skin. Original magnifica- tion, 63. C, Total protein extracts of SSc and control skin were probed with anti- VEGF165b–specific and anti- pan-VEGF antibodies. Results of a representative experiment are shown. SSc 1 to 3 indi- cates early SSc skin; SSc 4 and 5, late SSc skin. D, Densi- tometric analysis of blot bands. Data are meanSD of optical density in arbitrary units (a.u.). *P0.001 vs VEGF165b in control, **P0.001 vs pan-VEGF in control. E, Different concentra- tions of recombinant human (rh) VEGF165b were probed with anti-VEGF165b–specific and anti-pan-VEGF antibodies.

Article Snippet: This antibody has been shown using Western blotting and immunohistochemistry to specifically detect the VEGF165b isoform of VEGF, but not the pro-angiogenic VEGF165 isoform, even when present at 10,000-fold greater concentrations.9,12 Protein samples extracted from various human tissues, when probed with this antibody, revealed immunoblotting bands of the same size as that detected using a pan-VEGF antibody.9 Three commercially available anti-VEGF-A antibodies were used to investigate the expression of all isoforms of VEGF (including VEGF165b) – termed pan-VEGF: rabbit anti-human VEGF antibody A-20 and mouse anti-human VEGF antibody C-1 (both from Santa Cruz Biotechnology, Santa Cruz, CA, USA; cat. no. sc-152 and sc-7269, respectively), and mouse anti- human VEGF antibody (BD PharMingen, Heidelberg, Germany; cat. no. 555036).

Techniques: Expressing, Control, Recombinant

Journal: Circulation Research

Article Title: Overexpression of VEGF 165 b, an Inhibitory Splice Variant of Vascular Endothelial Growth Factor, Leads to Insufficient Angiogenesis in Patients With Systemic Sclerosis

doi: 10.1161/circresaha.111.242057

Figure Lengend Snippet: Figure 4. Circulating levels of VEGF165b and pan-VEGF. VEGF165b (A, B) and pan-VEGF (C, D) were mea- sured in SSc and control plasma by ELISA. Data are box plots with median and upper and lower quartiles. Proba- bility values were calculated by the nonparametric Mann-Whitney U test. A, #P0.001, *P0.002 vs control subjects. B, #P0.001, *P0.004 vs control subjects. C, #P0.001, *P0.002 vs control subjects. D, #P0.001, *P0.001 vs control subjects.

Article Snippet: This antibody has been shown using Western blotting and immunohistochemistry to specifically detect the VEGF165b isoform of VEGF, but not the pro-angiogenic VEGF165 isoform, even when present at 10,000-fold greater concentrations.9,12 Protein samples extracted from various human tissues, when probed with this antibody, revealed immunoblotting bands of the same size as that detected using a pan-VEGF antibody.9 Three commercially available anti-VEGF-A antibodies were used to investigate the expression of all isoforms of VEGF (including VEGF165b) – termed pan-VEGF: rabbit anti-human VEGF antibody A-20 and mouse anti-human VEGF antibody C-1 (both from Santa Cruz Biotechnology, Santa Cruz, CA, USA; cat. no. sc-152 and sc-7269, respectively), and mouse anti- human VEGF antibody (BD PharMingen, Heidelberg, Germany; cat. no. 555036).

Techniques: Control, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY