VEGF Polyclonal Antibody Catalog Number PA Product data sheet

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1 Website: thermofisher.com Customer Service (US): ext. 1 Technical Support (US): ext. 441 VEGF Polyclonal Antibody Catalog Number PA Product data sheet Details Size 500 µl Host/Isotope Class Type Immunogen Conjugate Form Concentration Purification Storage buffer Contains Storage Conditions Rabbit / IgG Polyclonal Antibody A synthetic peptide derived from N- terminal of human VEGF Unconjugated Liquid 0.2 mg/ml purified PBS, ph 7.6, with 0.2% BSA 15mM sodium azide 4 C Species Reactivity Species reactivity Published species Human, Mouse Tested Applications Dilution * Immunocytochemistry (ICC) 1:40-1:200 Immunofluorescence (IF) 1:40-1:200 Immunohistochemistry (Paraffin) (IHC (P)) Immunoprecipitation (IP) Western Blot (WB) Published Applications Western Blot (WB) Immunohistochemistry (Paraffin) (IHC (P)) Immunohistochemistry (IHC) Immunofluorescence (IF) Rat, Human, Mouse, Not Applicable 1:100 3 µg/ml 1 µg/ml See 3 publications below See 3 publications below See 5 publications below See 1 publications below * Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls. Product specific information PA targets Vascular Endothelial Growth Factor (VEGF) in IF, IHC (P), IP, and WB applications and shows reactivity with Human samples. The PA immunogen is a synthetic peptide derived from N-terminal of human VEGF. Background/Target Information VEGF belongs to a family of heparin binding growth factors and is the major mediator of vasculogenesis and angiogenesis. VEGF has a variety of effects on vascular endothelium, including the ability to promote endothelial cell viability, mitogenesis, chemotaxis, and vascular permeability. VEGF is a 45 kda homodimeric glycoprotein, but other isoforms can be produced through alternative splicing.

2 Product Images For VEGF Polyclonal Antibody Immunofluorescent analysis of Vascular Endothelial Growth Factor (VEGF, green) in Hela cells. The cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% Blocker BSA (Product # 37525) for 30 minutes at room temperature. Cells were stained with (left panel) or without (right panel) a VEGF polyclonal antibody (Product # PA ) at a dilution of 1:40 for at least 1 hour at room temperature, and then incubated with a Dylight 488 goat anti-rabbit IgG secondary antibody at a dilution of 1: 1000 for 45 minutes at room temperature. F-actin (both panels, red) was stained by Dylight 554 Phalloidin (Product # 21834) and nuclei (both panels, blue) were stained with DAPI (Product # 46190). Images were taken at 60X magnification. VEGF Antibody in Immunofluorescence (IF) Fig. 3. Increased expression of VEGF targets in the first trimester. (A) Heat map representing the relative expression and fold enrichment of genes repressed by VEGF (VEGFA) in the first ( n =4) and second ( n =3) trimester in OFT leaflets (created using Molecular Signatures Database v5.0). See B for color key. (B) GSEA of enrichment of VEGF repression from leaflets obtained from the first compared with the second trimester. The false discovery rate (FDR) q-value and normalized enrichment score (NES) are shown. (C-L) Immunofluorescence imaging of NFATC1 (red) and VEGF (green) protein expression patterns in early endocardial cushions and elongated leaflets. DAPI is in white. At the early cushion stage (C,D), fibrosa and ventricularis are not yet identifiable. E-H, the fibrosa layer; I-L, the ventricularis. Scale bar: 50 um. Immunohistochemistry analysis of VEGF showing staining in the cytoplasm of paraffin-embedded mouse kidney tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (ph 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddh2o and PBS, and then probed with a VEGF Rabbit Polyclonal Antibody (Product # PA ) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

3 VEGF Antibody in Immunohistochemistry (IHC) Published figure using VEGF polyclonal antibody (Product # PA ) in Immunohistochemistry Immunohistochemistry analysis of VEGF showing staining in the cytoplasm of paraffin-embedded human liver tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (ph 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddh2o and PBS, and then probed with a VEGF Rabbit Polyclonal Antibody (Product # PA ) diluted in 3% BSA-PBS at a dilution of 1:100 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting. VEGF Antibody in Immunohistochemistry (IHC) Published figure using VEGF polyclonal antibody (Product # PA ) in Immunohistochemistry Formalin-fixed, paraffin-embedded human angiosarcoma stained with VEGF antibody using peroxidaseconjugate and AEC. Note cytoplasmic staining of tumor cells.

4 Western blot analysis of Vascular Endothelial Growth Factor (VEGF) was performed by loading 20ug of MCF7, HepG2, HeLa, A431, A549 whole cell lysates and 10uL of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. VEGF was detected at ~45kD and ~24kD using an VEGF polyclonal antibody (Product # PA ) using a concentration of 1 µg/ml in 5% milk in TBST overnight at 4C on a rocking platform, followed by a goat anti-rabbit IgG-HRP secondary antibody (Product # 31460) at a dilution of 1:20,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura substrate (Product # 34076). Western blot analysis of Vascular Endothelial Growth Factor (VEGF) was performed by loading 1ug of indicated recombinant VEGF isoforms, and 10ul of PageRuler PlusPrestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. VEGF was detected at 14kD, 19kD and kd using a VEGF polyclonal antibody (Product # PA ) at a dilution of 1:500 in blocking buffer overnight at 4C on a rocking platform, followed by an HRPconjugated goat anti-rabbit IgG (Fc) secondary antibody (Product # 31463) at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

5 Western blot of Vascular Endothelial Growth Factor using Vascular Endothelial Growth Factor Polyclonal Antibody (Product # PA ) on VEGF Cells. Immunoprecipitation of Vascular Endothelial Growth Factor (VEGF) was performed on HeLa cell lysate. Antigen-antibody complexes were formed by incubating 750ug of whole cell lysate with 3ug of a VEGF polyclonal antibody (Product # PA ) overnight on a rocking platform at 4C. The immune complexes were captured on 50ul Protein A/G Plus Agarose (Product # 20423), washed extensively, and eluted with Lane Marker Reducing Sample Buffer (Product # HeLa cell lysate 20ug was loaded as a positive control (left lane). Samples were resolved on a 4-20% Tris-Glycing Polyacrylamide gel, transferred to a Nitrocellulose membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a VEGF polyclonal antibody (Product # PA ) at a concentration of 1 µg/ml in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG (Fc) secondary antibody (Product # 31463) at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

6 PubMed References For VEGF Polyclonal Antibody 3 Western Blot References PA was used in western blot to suggest that mir-24 is a central regulator of race-related prostate cancer Human / 1:200 Mouse / 1:500 Oncotarget (Mar 2017; 8: 16581) "The role of mir-24 as a race related genetic factor in prostate cancer." Author(s):Hashimoto Y,Shiina M,Kato T,Yamamura S,Tanaka Y,Majid S,Saini S,Shahryari V,Kulkarni P,Dasgupta P,Mitsui Y, Sumida M,Deng G,Tabatabai L,Kumar D,Dahiya R PubMed Article URL: PA was used in western blot to study the roles of angiocrine secretion and accumulation of HIF-1alpha in the mechanism by which phosphorylation of endothelial cell profilin-1 promotes the progression of glioblastoma Nature cell biology (May 2014; 16: 445) "Profilin-1 phosphorylation directs angiocrine expression and glioblastoma progression through HIF-1 accumulation." Author(s):Fan Y,Potdar AA,Gong Y,Eswarappa SM,Donnola S,Lathia JD,Hambardzumyan D,Rich JN,Fox PL PubMed Article URL: PA was used in western blot to study the role of COX-2 in the upregulation of CNS laminin in a murine model of ischemic injury Mouse / 1:200 3 Immunohistochemistry (Paraffin) References Journal of neuroinflammation (Jul 2012; 9: null) "Inflammation modulates expression of laminin in the central nervous system following ischemic injury." Author(s):Ji K,Tsirka SE PubMed Article URL: PA was used in immunohistochemistry - paraffin section to characterize the histopathologic prognosis of renal cell carcinoma by combined use of EGF and COX-1 immunohistochemistry Not Applicable / 1:200 Not Applicable / 1:200 Not Applicable / 1:100 International journal of clinical and experimental pathology (Jun 2016; 8: 8165) "Combined use of COX-1 and VEGF immunohistochemistry refines the histopathologic prognosis of renal cell carcinoma." Author(s):Osman WM,Youssef NS PubMed Article URL: PA was used in immunohistochemistry - paraffin section to study the attenuation of VEGF expression and enhancement of necrosis in mammary carcinoma by deletion of IL-33R Oncotarget (Apr 2016; 7: 18106) "Deletion of IL-33R attenuates VEGF expression and enhances necrosis in mammary carcinoma." Author(s):Milosavljevic MZ,Jovanovic IP,Pejnovic NN,Mitrovic SL,Arsenijevic NN,Simovic Markovic BJ,Lukic ML PubMed Article URL: PA was used in immunohistochemistry - paraffin section to characterize human cardiac valve development by endocardial-to-mesenchymal transformation and mesenchymal cell colonization Not Applicable / 1:4000 Not Applicable / 1: Immunohistochemistry References Development (Cambridge, England) (Feb 2016; 143: 473) "Endocardial-to-mesenchymal transformation and mesenchymal cell colonization at the onset of human cardiac valve development." Author(s):Monaghan MG,Linneweh M,Liebscher S,Van Handel B,Layland SL,Schenke-Layland K PubMed Article URL: PA was used in immunohistochemistry - paraffin section to characterize the histopathologic prognosis of renal cell carcinoma by combined use of EGF and COX-1 immunohistochemistry International journal of clinical and experimental pathology (Jun 2016; 8: 8165) "Combined use of COX-1 and VEGF immunohistochemistry refines the histopathologic prognosis of renal cell carcinoma." Author(s):Osman WM,Youssef NS PubMed Article URL:

7 PA was used in immunohistochemistry to study the rmechanism by which tumor endothelial FasL promotes tumor immune tolerance Human / 1:100 Rat / 1:100 Rat / 1:100 Human / Not Cited Nature medicine (Jun 2014; 20: 607) "Tumor endothelium FasL establishes a selective immune barrier promoting tolerance in tumors." Author(s):Motz GT,Santoro SP,Wang LP,Garrabrant T,Lastra RR,Hagemann IS,Lal P,Feldman MD,Benencia F,Coukos G PubMed Article URL: PA was used in immunohistochemistry to study age-related deterioration of the optic nerve in a rat model Age (Dordrecht, Netherlands) (Apr 2014; 36: 519) "Aging-related changes of optic nerve of Wistar albino rats." Author(s):El-Sayyad HI,Khalifa SA,El-Sayyad FI,Al-Gebaly AS,El-Mansy AA,Mohammed EA PubMed Article URL: PA was used in immunohistochemistry to study the patterns of proliferation and maturation of microvessels in arteriovenous malformations Journal of cutaneous pathology (Jun 2012; 39: 610) "Proliferation and maturation of microvessels in arteriovenous malformations--expression patterns of angiogenic and cell cycle-dependent factors." Author(s):Meijer-Jorna LB,van der Loos CM,Teeling P,de Boer OJ,Florquin S,van der Horst CM,van der Wal AC PubMed Article URL: PA was used in immunohistochemistry to study blood vessel stabilization and the expression of pro-angiogenic VEGF and Ang-2 in castration-resistant prostate cancer Human / 1:2 1 Immunofluorescence References The Prostate (May 2012; 72: 705) "Castration resistant prostate cancer is associated with increased blood vessel stabilization and elevated levels of VEGF and Ang-2." Author(s):Tomi TT,Gustavsson H,Wang W,Jennbacken K,Welén K,Damber JE PubMed Article URL: PA was used in immunohistochemistry - paraffin section to characterize human cardiac valve development by endocardial-to-mesenchymal transformation and mesenchymal cell colonization Development (Cambridge, England) (Feb 2016; 143: 473) "Endocardial-to-mesenchymal transformation and mesenchymal cell colonization at the onset of human cardiac valve development." Author(s):Monaghan MG,Linneweh M,Liebscher S,Van Handel B,Layland SL,Schenke-Layland K PubMed Article URL:

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