Human VEGFR2/KDR/Flk-1 Antibody

AF357 28 Citations Datasheet / COA / SDS

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AF357-SP

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Citations (28)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human VEGFR2/KDR/Flk-1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human VEGFR2/KDR/Flk‑1 in direct ELISAs and Western blots. In direct ELISAs, approximately 40% cross-reactivity with recombinant mouse VEGFR2 is observed and less than 10% cross-reactivity with recombinant human (rh) VEGFR1 and rhVEGFR3 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant human VEGFR2/KDR/Flk-1
Ala20-Glu764
Accession # AAC16450

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Neutralization

Measured by its ability to neutralize VEGFR2/KDR/Flk‑1-mediated inhibition of proliferation in HUVEC human umbilical vein endothelial cells. The Neutralization Dose (ND₅₀) is typically 0.05-0.25 µg/mL in the presence of 30 ng/mL Recombinant Human VEGFR2/KDR/Flk‑1 Fc Chimera and 5 ng/mL Recombinant Human VEGF₁₆₅.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot

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Detection of Human VEGFR2/KDR/Flk‑1 by Western Blot. Western blot shows lysate of HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human VEGFR2/KDR/Flk-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF357) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for VEGFR2/KDR/Flk-1 at approximately 200-250 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

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VEGFR2/KDR/Flk‑1 in Human Placenta. VEGFR2/KDR/Flk-1 was detected in immersion fixed paraffin-embedded sections of human placenta using 15 µg/mL Goat Anti-Human VEGFR2/KDR/Flk-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF357) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-AEC Cell & Tissue Staining Kit (red; Catalog # CTS009) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Immunohistochemistry

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VEGFR2/KDR/Flk‑1 in Human Kidney. VEGFR2/KDR/Flk-1 was detected in immersion fixed paraffin-embedded sections of human kidney using Goat Anti-Human VEGFR2/KDR/Flk-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF357) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Neutralization

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VEGFR2/KDR/Flk‑1 Inhibi-tion of VEGF-dependent Cell Proliferation and Neutral-ization by Human VEGFR2/KDR/Flk‑1 Anti-body. Recombi-nant Human VEGFR2/KDR/Flk-1 Fc Chi-mera (Catalog # 357-KD) inhibits Recombinant Human VEGF₁₆₅(Catalog # 293-VE) induced proliferation in HUVEC human umbilical vein endothelial cells in a dose-dependent manner (orange line). Inhibition of Recombinant Human VEGF₁₆₅(5 ng/mL) activity elicited by Recombinant Human VEGFR2/KDR/Flk-1 Fc Chi-mera (30 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human VEGFR2/KDR/Flk-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF357). The ND₅₀is typically 0.05-0.25 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & 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.

Background: VEGFR2/KDR/Flk-1

VEGFR2 (KDR/Flk-1), VEGFR1 (Flt-1) and VEGFR3 (Flt-4) belong to the class III subfamily of receptor tyrosine kinases (RTKs). All three receptors contain seven immunoglobulin-like repeats in their extracellular domains and kinase insert domains in their intracellular regions. The expression of VEGFR1, 2, and 3 is almost exclusively restricted to the endothelial cells. These receptors are likely to play essential roles in vasculogenesis and angiogenesis.

VEGFR2 cDNA encodes a 1356 amino acid (aa) residue precursor protein with a 19 aa residue signal peptide. Mature VEGFR2 is composed of a 745 aa residue extracellular domain, a 25 aa residue transmembrane domain and a 567 aa residue cytoplasmic domain. In contrast to VEGFR1 which binds both PlGF and VEGF with high affinity, VEGFR2 binds VEGF but not PlGF with high affinity. The recombinant soluble VEGFR2/Fc chimera binds VEGF with high affinity and is a potent VEGF antagonist.

References

Ferra, N. and R. Davis-Smyth (1997) Endocrine Reviews 18:4.

Long Name

Vascular Endothelial Growth Factor Receptor 2

Entrez Gene IDs

3791 (Human); 16542 (Mouse)

Alternate Names

CD309 antigen; CD309; EC 2.7.10; EC 2.7.10.1; Fetal liver kinase 1; fetal liver kinase-1; Flk1; Flk-1; FLK1tyrosine kinase growth factor receptor; KDR; kinase insert domain receptor (a type III receptor tyrosine kinase); Kinase insert domain receptor; KRD1; Ly73; Protein-tyrosine kinase receptor flk-1; soluble VEGFR2; vascular endothelial growth factor receptor 2; VEGF R2; VEGFR; VEGFR2; VEGFR-2

Product Datasheets

Product Datasheet

COA

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Related Research Areas

Citations for Human VEGFR2/KDR/Flk-1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

28 Citations: Showing 1 - 10
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Atypical chemokine receptor ACKR3/CXCR7 controls postnatal vasculogenesis and arterial specification by mesenchymal stem cells via Notch signaling
Authors: ST Wei, YC Huang, ML Hsieh, YJ Lin, WC Shyu, HC Chen, CH Hsieh
Cell Death Dis, 2020;11(5):307.
Species: Human
Sample Types: Cell Culture Supernates
Applications: Western Blot
ATF-2 and Tpl2 regulation of endothelial cell cycle progression and apoptosis
Authors: GW Fearnley, AM Latham, M Hollstein, AF Odell, S Ponnambala
Cell. Signal., 2019;66(0):109481.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
Targeting chemoresistant colorectal cancer via systemic administration of a BMP7 variant
Authors: V Veschi, LR Mangiapane, A Nicotra, S Di Franco, E Scavo, T Apuzzo, DS Sardina, M Fiori, A Benfante, ML Colorito, G Cocorullo, F Giuliante, C Cipolla, G Pistone, MR Bongiorno, A Rizzo, CM Tate, X Wu, S Rowlinson, LF Stancato, M Todaro, R De Maria, G Stassi
Oncogene, 2019;0(0):.
Species: Xenograft
Sample Types: Whole Tissue
Applications: IHC-P
Endophilin-A2 dependent VEGFR2 endocytosis promotes sprouting angiogenesis
Authors: G Genet, K Boyé, T Mathivet, R Ola, F Zhang, A Dubrac, J Li, N Genet, L Henrique G, L Benedetti, S Künzel, L Pibouin-Fr, JL Thomas, A Eichmann
Nat Commun, 2019;10(1):2350.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
Endoglin prevents vascular malformation by regulating flow-induced cell migration and specification through VEGFR2 signalling
Authors: Y Jin, L Muhl, M Burmakin, Y Wang, AC Duchez, C Betsholtz, HM Arthur, L Jakobsson
Nat. Cell Biol., 2017;0(0):.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC
Lymphatic endothelial progenitors originate from plastic myeloid cells activated by toll-like receptor-4
Authors: LD Volk-Drape, KL Hall, AC Wilber, S Ran
PLoS ONE, 2017;12(6):e0179257.
Species: Human
Sample Types: Whole Cells
Applications: ICC
Affimer proteins are versatile and renewable affinity reagents
Authors: C Tiede, R Bedford, SJ Heseltine, G Smith, I Wijetunga, R Ross, D AlQallaf, AP Roberts, A Balls, A Curd, RE Hughes, H Martin, SR Needham, LC Zanetti-Do, Y Sadigh, TP Peacock, AA Tang, N Gibson, H Kyle, GW Platt, N Ingram, T Taylor, LP Coletta, I Manfield, M Knowles, S Bell, F Esteves, A Maqbool, RK Prasad, M Drinkhill, RS Bon, V Patel, SA Goodchild, M Martin-Fer, RJ Owens, JE Nettleship, ME Webb, M Harrison, JD Lippiat, S Ponnambala, M Peckham, A Smith, PK Ferrigno, M Johnson, MJ McPherson, DC Tomlinson
Elife, 2017;6(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
VEGFR3 Modulates Vascular Permeability by Controlling VEGF/VEGFR2 Signaling
Authors: K Heinolaine, S Karaman, G D'Amico, T Tammela, R Sormunen, L Eklund, K Alitalo, G Zarkada
Circ. Res, 2017;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
VEGF induces signalling and angiogenesis by directing VEGFR2 internalisation via macropinocytosis
J Cell Sci, 2016;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization
Differential sensitivity of prostate tumor derived endothelial cells to sorafenib and sunitinib.
Authors: Fiorio Pla, Alessand, Brossa, Alessia, Bernardini, Michela, Genova, Tullio, Grolez, Guillaum, Villers, Arnaud, Leroy, Xavier, Prevarskaya, Natalia, Gkika, Dimitra, Bussolati, Benedett
BMC Cancer, 2014;14(0):939.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

VEGF-A isoforms differentially regulate ATF-2-dependent VCAM-1 gene expression and endothelial-leukocyte interactions.
Authors: Fearnley G, Odell A, Latham A, Mughal N, Bruns A, Burgoyne N, Homer-Vanniasinkam S, Zachary I, Hollstein M, Wheatcroft S, Ponnambalam S
Mol Biol Cell, 2014;25(16):2509-21.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Sustained delivery of VEGF maintains innervation and promotes reperfusion in ischemic skeletal muscles via NGF/GDNF signaling.
Authors: Shvartsman, Dmitry, Storrie-White, Hannah, Lee, Kangwon, Kearney, Cathal, Brudno, Yevgeny, Ho, Nhi, Cezar, Christin, McCann, Corey, Anderson, Erin, Koullias, John, Tapia, Juan Car, Vandenburgh, Herman, Lichtman, Jeff W, Mooney, David J
Mol Ther, 2014;22(7):1243-53.
Species: Human
Sample Types: Whole Cells
Applications: Functional Assay

Control of angiogenesis by galectins involves the release of platelet-derived proangiogenic factors.
Authors: Etulain J, Negrotto S, Tribulatti M, Croci D, Carabelli J, Campetella O, Rabinovich G, Schattner M
PLoS ONE, 2014;9(4):e96402.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Endothelial cell adhesion to soluble vascular endothelial growth factor receptor-1 triggers a cell dynamic and angiogenic phenotype.
Authors: Orecchia A, Mettouchi A, Uva P, Simon G, Arcelli D, Avitabile S, Ragone G, Meneguzzi G, Pfenninger K, Zambruno G, Failla C
FASEB J, 2014;28(2):692-704.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

MicroRNA-24 suppression of N-deacetylase/N-sulfotransferase-1 (NDST1) reduces endothelial cell responsiveness to vascular endothelial growth factor A (VEGFA).
Authors: Kasza Z, Fredlund Fuchs P, Tamm C, Eriksson A, O'Callaghan P, Heindryckx F, Spillmann D, Larsson E, Le Jan S, Eriksson I, Gerwins P, Kjellen L, Kreuger J
J Biol Chem, 2013;288(36):25956-63.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

KDR identifies a conserved human and murine hepatic progenitor and instructs early liver development.
Authors: Goldman O, Han S, Sourisseau M, Dziedzic N, Hamou W, Corneo B, D'Souza S, Sato T, Kotton D, Bissig K, Kalir T, Jacobs A, Evans T, Evans M, Gouon-Evans V
Cell Stem Cell, 2013;12(6):748-60.
Species: Human
Sample Types: Whole Cells
Applications: ICC

Essential role for endocytosis in the growth factor-stimulated activation of ERK1/2 in endothelial cells.
Authors: Gourlaouen, Morgane, Welti, Jonathan, Vasudev, Naveen S, Reynolds, Andrew R
J Biol Chem, 2013;288(11):7467-80.
Species: Human
Sample Types: Whole Cells
Applications: ICC

Differential expression of Vegfr-2 and its soluble form in preeclampsia.
Authors: Munaut C, Lorquet S, Pequeux C, Coulon C, Le Goarant J, Chantraine F, Noel A, Goffin F, Tsatsaris V, Subtil D, Foidart JM
PLoS ONE, 2012;7(3):e33475.
Species: Human
Sample Types: Plasma
Applications: Western Blot

Vascular endothelial growth factor drives autocrine epithelial cell proliferation and survival in chronic rhinosinusitis with nasal polyposis.
Authors: Lee HS, Myers A, Kim J
Am. J. Respir. Crit. Care Med., 2009;180(11):1056-67.
Species: Human
Sample Types: Whole Tissue
Applications: IHC

The Down syndrome critical region gene 1 short variant promoters direct vascular bed-specific gene expression during inflammation in mice.
Authors: Minami T, Yano K, Miura M, Kobayashi M, Suehiro J, Reid PC, Hamakubo T, Ryeom S, Aird WC, Kodama T
J. Clin. Invest., 2009;119(8):2257-70.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Enhancement of angiogenic effectors through hypoxia-inducible factor in preterm primate lung in vivo.
Authors: Asikainen TM, Waleh NS, Schneider BK, Clyman RI, White CW
Am. J. Physiol. Lung Cell Mol. Physiol., 2006;291(4):L588-95.
Species: Primate - Papio anubis (Olive Baboon)
Sample Types: Tissue Homogenates
Applications: Western Blot

Placenta growth factor in diabetic wound healing: altered expression and therapeutic potential.
Authors: Cianfarani F, Zambruno G, Brogelli L, Sera F, Lacal PM, Pesce M, Capogrossi MC, Failla CM, Napolitano M, Odorisio T
Am. J. Pathol., 2006;169(4):1167-82.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Activation of hypoxia-inducible factors in hyperoxia through prolyl 4-hydroxylase blockade in cells and explants of primate lung.
Authors: Asikainen TM, Schneider BK, Waleh NS, Clyman RI, Ho WB, Flippin LA, Gunzler V, White CW
Proc. Natl. Acad. Sci. U.S.A., 2005;102(29):10212-7.
Species: Human
Sample Types: Whole Cells
Applications: ICC

VEGF164(165) as the pathological isoform: differential leukocyte and endothelial responses through VEGFR1 and VEGFR2.
Authors: Usui T, Ishida S, Yamashiro K, Kaji Y, Poulaki V, Moore J, Moore T, Amano S, Horikawa Y, Dartt D, Golding M, Shima DT, Adamis AP
Invest. Ophthalmol. Vis. Sci., 2004;45(2):368-74.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

PEDF derived from glial Muller cells: a possible regulator of retinal angiogenesis.
Authors: Eichler W, Yafai Y, Keller T, Wiedemann P, Reichenbach A
Exp. Cell Res., 2004;299(1):68-78.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Proangiogenic properties of human myeloma cells: production of angiopoietin-1 and its potential relationship to myeloma-induced angiogenesis.
Authors: Giuliani N, Colla S, Lazzaretti M, Sala R, Roti G, Mancini C, Bonomini S, Lunghi P, Hojden M, Genestreti G, Svaldi M, Coser P, Fattori PP, Sammarelli G, Gazzola GC, Almici C, Caramatti C, Mangoni L, Rizzoli V
Blood, 2003;102(2):638-45.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Malignant mesothelioma growth inhibition by agents that target the VEGF and VEGF-C autocrine loops.
Authors: Masood R, Kundra A, Zhu S, Xia G, Scalia P, Smith DL, Gill PS
Int. J. Cancer, 2003;104(5):603-10.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Human herpesvirus 8 (HHV-8)-encoded cytokines induce expression of and autocrine signaling by vascular endothelial growth factor (VEGF) in HHV-8-infected primary-effusion lymphoma cell lines and mediate VEGF-independent antiapoptotic effects.
Authors: Okruzhnov Y, Nicholas J
J. Virol., 2001;75(22):10933-40.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot