VEGF Antibody (56-1) [Unconjugated] - 165b

VEGF165bwas detected in immersion fixed paraffin-embedded sections of human placenta using Mouse Anti-Human VEGF165bMonoclonal Antibody (Catalog # MAB3045) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to syncytiotrophoblasts. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

VEGF expression determined by Western blot and immunoprecipitation.A. Western blot using LiCor Odyssey to simultaneously image pan-VEGF and VEGF-A165b probed western blot. Two different podocyte samples, and a primary RPE sample were run on a gel and probed with antibodies to VEGF-A165b (mouse monoclonal anti-CTRSLTRKD, and 680nm-donkey anti-mouse, top image) and pan-VEGF (rabbit polyclonal anti-VEGF, and 800nm-donkey anti-rabbit, middle image). The bottom image is the pseudocoloured combined image (600nm green, 800nm red). Note the red VEGF165, but yellow VEGF-A165b. MWM = molecular weight marker. d = dimer, m = monomer. B. Protein extracted from human cell lines (adenoma and adenocarcinoma(AC)) subjected to immunoprecipitation (IP) for VEGF-A165b and immunoblotting (IB) for total VEGF-A. A clear strong band was seen in the IP for both cell types at ∼23kDa and ∼46kDa, consistent with the IP for recombinant human VEGF-A165b. A weaker band was seen in the input protein (not subjected to IP), and a second band slightly higher in the AC. A weak band at approximately 56kDa and 28kDa was seen in all lanes subjected to IP, including the VEGF-A165a band, but not seen in the recombinant human VEGF-A165b not subjected to IP, indicating that this is cross reactivity with the IgG. This band was clearly above the VEGF-A165b bands. C. Protein extracted from human cell lines (adenoma and adenocarcinoma(AC)) subjected to immunoprecipitation (IP) for VEGF-A and immunoblotting (IB) for VEGF-A165b. A clear strong band was seen in the IP for both cell types at ∼23kDa, the same size as recombinant human VEGF-A165b. In the input a band at ∼46Da was seen predominantly, for both cell types, labelled as VEGF-A165b dimers. D. Mouse tissues probed with VEGF-A165b antibody detect mouse IgG due to the secondary antibody. Top image, western blot of mouse tissues, recombinant mouse IgG or human VEGF-A165b or VEGF-A165b probed with mouse anti-CTRSLTRKD, and 680nm-donkey anti-mouse IgG. Bottom image blot of same tissues, probed without primary antibody. The same bands are seen in the mouse tissues. Spl = spleen, Col = colon, Hrt = heart, Lng = lung, Liv = liver, Kid = kidney. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23935865), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IHC, ICC/IF
• Clone: 56-1
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

VEGF Antibody (56-1) [Unconjugated] - 165b Summary

• Immunogen: KLH-conjugated human VEGF_165b synthetic peptide
  TCRSLTRKD
  Accession # AAL27435
• Specificity: Detects human VEGF_165b in direct ELISAs and Western blots. Recognizes an epitope within a 9 amino acid sequence at the C-terminus of human VEGF_165b. In direct ELISAs and Western blots, no cross-reactivity with recombinant human (rh) VEGF₂₀₆, rhVEGF-B₁₆₇, rhVEGF-B₁₈₆, rhVEGF-C, or rhVEGF-D is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: VEGFA
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunofluorescence
  • Immunohistochemistry 8-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for VEGF Antibody (56-1) [Unconjugated] - 165b

• MVCD1
• VAS
• vascular endothelial growth factor A
• Vascular permeability factor
• Vasculotropin
• VEGF
• VEGFA
• VEGF-A
• VEGFMGC70609
• VPF
• VPFvascular endothelial growth factor

Background

VEGF_165b is the protein resulting from an mRNA splice variant of VEGF. Alternative exon usage results in a 165 amino acid product that is identical to VEGF₁₆₅ except for 6 unique amino acids at the C-terminus. VEGF₁₆₅b inhibits proliferation and migration of endothelial cells and is down-regulated in renal cell carcinomas.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for VEGF Antibody (MAB3045)(24)

Publications using MAB3045

• Chen, M;Chen, J;Liu, Y;Wang, X;Yao, M;Chen, J;Zhang, J;Huang, Q; Senescent Macrophages Promote Age-Related Revascularization Impairment by Increasing Antiangiogenic VEGF-A165B Expression Aging cell 2025-04-17 [PMID: 40243169] (Western Blot, Neutralization, Mouse)
• Quan, B;Li, Z;Yang, H;Li, S;Yan, X;Wang, Y; The splicing factor YBX1 promotes the progression of osteosarcoma by upregulating VEGF165 and downregulating VEGF165b Heliyon 2023-08-01 [PMID: 37554848] (Western Blot, Human)
• T Jia, T Jacquet, F Dalonneau, P Coudert, E Vaganay, C Exbrayat-H, J Vollaire, V Josserand, F Ruggiero, JL Coll, B Eymin FGF-2 promotes angiogenesis through a SRSF1/SRSF3/SRPK1-dependent axis that controls VEGFR1 splicing in endothelial cells Bmc Biology, 2021-08-25;19(1):173. 2021-08-25 [PMID: 34433435] (Western Blot, Human)
• A Prodjohard, AN Vidyanti, NA Susianti, Sudarmanta, S Sutarni, I Setyoprano Higher level of acute serum VEGF and larger infarct volume are more frequently associated with post-stroke cognitive impairment PLoS ONE, 2020-10-05;15(10):e0239370. 10/5/2020 [PMID: 33017430]
• Lindsey G. Hawke, Mara K. M. Whitford, Mark L. Ormiston The Production of Pro-angiogenic VEGF-A Isoforms by Hypoxic Human NK Cells Is Independent of Their TGF-beta -Mediated Conversion to an ILC1-Like Phenotype Frontiers in Immunology 2020-08-25 [PMID: 32983113]
• AF Stassi, F Gasser, MML Velázquez, EM Belotti, NC Gareis, F Rey, HH Ortega, NR Salvetti, ME Baravalle Contribution of the VEGF system to the follicular persistence associated with bovine cystic ovaries Theriogenology, 2019-07-03;138(0):52-65. 2019-07-03 [PMID: 31301448] (IHC, IHC-P, Western Blot, Bovine, Mouse)
• Patrícia Matos Biselli-Chicote, Joice Matos Biselli, Bianca R Cunha, Rodrigo Castro, José Victor Maniglia, Dalísio de Santi Neto et al. Overexpression of Antiangiogenic Vascular Endothelial Growth Factor Isoform and Splicing Regulatory Factors in Oral, Laryngeal and Pharyngeal Squamous Cell Carcinomas Asian Pac. J. Cancer Prev 2017-08-28 [PMID: 28843252]
• Shakun Karki, Doan T. M. Ngo, Melissa G. Farb, Song Young Park, Samantha M. Saggese, Naomi M. Hamburg et al. WNT5A regulates adipose tissue angiogenesis via antiangiogenic VEGF-A165b in obese humans American Journal of Physiology-Heart and Circulatory Physiology 2017-07-01 [PMID: 28411232]
• MASAHIRO NAGASAKI, SEIJI KONDO, YOSHIKI MUKUDAI, TAKAAKI KAMATANI, AYAKO AKIZUKI, ATSUSHI YASO et al. Clinicopathological implications of vascular endothelial growth factor 165b expression in oral squamous cell carcinoma stroma Oncology Reports 2016-07-01 [PMID: 27221145]
• Kim H, Kim K, Kim B, Cho M, Lee S The effect of vascular endothelial growth factor on osteoclastogenesis in rheumatoid arthritis. PLoS ONE, 2015-04-20;10(4):e0124909. 2015-04-20 [PMID: 25894998] (IHC, Human)
• James G. Carter, Melissa V. R. Gammons, Gopinath Damodaran, Amanda J. Churchill, Steven J. Harper, David O. Bates The carboxyl terminus of VEGF-A is a potential target for anti-angiogenic therapy Angiogenesis 2015-01-01 [PMID: 25274272] (ELISA Capture, N/A)
• Bills V, Hamdollah-Zadeh M, Soothill P, Harper S, Bates D The role of VEGF-A165b in trophoblast survival. BMC Pregnancy Childbirth, 2014-08-15;14(0):278. 2014-08-15 [PMID: 25128406] (Neutralization, Human)
• Doan T.M. Ngo, Melissa G. Farb, Ryosuke Kikuchi, Shakun Karki, Stephanie Tiwari, Sherman J. Bigornia et al. Antiangiogenic Actions of Vascular Endothelial Growth Factor-A 165 b, an Inhibitory Isoform of Vascular Endothelial Growth Factor-A, in Human Obesity Circulation 2014-09-23 [PMID: 25116954]
• Van't Hull E, Bron S, Henry L, Ifticene-Treboux A, Turrini R, Coukos G, Delaloye J, Doucey M Bone marrow-derived cells are implicated as a source of lymphatic endothelial progenitors in human breast cancer. Oncoimmunology, 2014-06-25;3(0):e29080. 2014-06-25 [PMID: 25101222] (Flow Cytometry, Human)
• Bates , David O, Mavrou , Athina, Qiu , Yan, Carter , James G, Hamdollah-Zadeh , Maryam, Barratt , Shaney, Gammons , Melissa, Millar , Ann B, Salmon , Andrew H, Oltean , Sebastia, Harper , Steven J Detection of VEGF-A(xxx)b isoforms in human tissues. PLoS ONE, 2013-07-31;8(7):e68399. 2013-07-31 [PMID: 23935865] (Western Blot, Human)
• Parul Gupta, Chris Adkins, Paul Lockman, Sanjay K. Srivastava Metastasis of Breast Tumor Cells to Brain Is Suppressed by Phenethyl Isothiocyanate in a Novel In Vivo Metastasis Model PLoS ONE 2013-06-27 [PMID: 23826254]
• Jeanette Woolard, William Vousden, Steven J. Moss, Arjun Krishnakumar, Melissa Vr Gammons, David G Nowak et al. Borrelidin modulates the alternative splicing of VEGF in favour of anti-angiogenic isoforms Chem. Sci. 2011-02-01 [PMID: 22822423]
• Pavel Makarevich, Zoya Tsokolaeva, Alexander Shevelev, Igor Rybalkin, Evgeny Shevchenko, Irina Beloglazova et al. Combined Transfer of Human VEGF165 and HGF Genes Renders Potent Angiogenic Effect in Ischemic Skeletal Muscle PLoS ONE 2012-06-13 [PMID: 22719942]
• Merdzhanova G, Gout S, Keramidas M, Edmond V, Coll JL, Brambilla C, Brambilla E, Gazzeri S, Eymin B The transcription factor E2F1 and the SR protein SC35 control the ratio of pro-angiogenic versus antiangiogenic isoforms of vascular endothelial growth factor-A to inhibit neovascularization in vivo. Oncogene, 2010-07-19;29(39):5392-403. 2010-07-19 [PMID: 20639906] (Western Blot, Human)
• Jing Hua, Christine Spee, Satoru Kase, Emma S. Rennel, Anette L. Magnussen, Yan Qiu et al. Recombinant Human VEGF165b Inhibits Experimental Choroidal Neovascularization Investigative Opthalmology & Visual Science 2010-08-01 [PMID: 20237252] (Western Blot, Human)
• Anette L. Magnussen, Emma S. Rennel, Jing Hua, Heather S. Bevan, Nicholas Beazley Beazley Long, Christina Lehrling et al. VEGF-A165b Is Cytoprotective and Antiangiogenic in the Retina Investigative Opthalmology & Visual Science 2010-08-01 [PMID: 20237249]
• Varet J, Varet , Julia, Douglas SK, Douglas , Samantha, Gilmartin L, Gilmartin , Laura, Medford , Andrew R, Bates , David O, Harper , Steven J, Millar , Ann B VEGF in the lung: a role for novel isoforms. Am J Physiol Lung Cell Mol Physiol, 2010-03-12;298(6):L768-74. 2010-03-12 [PMID: 20228180] (ELISA Development, IHC-P, Western Blot, Human)
• Rennel ES, Varey AH, Churchill AJ, Wheatley ER, Stewart L, Mather S, Bates DO, Harper SJ VEGF(121)b, a new member of the VEGF(xxx)b family of VEGF-A splice isoforms, inhibits neovascularisation and tumour growth in vivo. Br. J. Cancer, 2009-08-25;101(7):1183-93. 2009-08-25 [PMID: 19707198] (Western Blot, Human)
• Bates DO, MacMillan PP, Manjaly JG, Qiu Y, Hudson SJ, Bevan HS, Hunter AJ, Soothill PW, Read M, Donaldson LF, Harper SJ The endogenous anti-angiogenic family of splice variants of VEGF, VEGFxxxb, are down-regulated in pre-eclamptic placentae at term. Clin. Sci., 2006-05-01;110(5):575-85. 2006-05-01 [PMID: 16451124] (ELISA Development, IHC-P, Western Blot, Human)

FAQs for VEGF Antibody (MAB3045). (Showing 1 - 1 of 1 FAQs).

1. Why is the molecular weight of VEGF different from the similar antibody, for some companies the the molecular weight is 40KD)?
   • I can't comment on another company's antibody because I don't have any information about their products. I can tell you that VEGF is expressed in a variety of isoforms and is subject to various post-translational modifications that influence its apparent molecular weight in an SDS-PAGE gel compared to the theoretical molecular weight.

Bioinformatics

• Gene Symbol: VEGFA
• Uniprot:
  • AAL27435()