Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Primate - Papio anubis (Olive Baboon), Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Immunocytochemistry, Immunoprecipitation, ELISA Capture, FACS, Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all VEGFR2/KDR/Flk-1 Primary Antibodies »

Product Specifications

Immunogen

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

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

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

Scientific Data Images for Human VEGFR2/KDR/Flk-1 Antibody

Detection of Human VEGFR2/KDR/Flk-1 antibody by Western Blot.

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.
VEGFR2/KDR/Flk-1 antibody in Human Placenta by Immunohistochemistry (IHC-P).

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.
VEGFR2/KDR/Flk-1 antibody in Human Kidney by Immunohistochemistry (IHC-P).

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.
VEGFR2/KDR/Flk‑1 Inhibi-tion of VEGF-dependent Cell Proliferation and Neutral-ization by Human VEGFR2/KDR/Flk‑1 Anti-body.

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 ND50 is typically 0.05-0.25 µg/mL.
Detection of Human VEGFR2/KDR/Flk-1 by Immunohistochemistry

Detection of Human VEGFR2/KDR/Flk-1 by Immunohistochemistry

BMP7v exerts antiangiogenic effects and sensitizes chemoresistant CSCs to standard therapy. a Azan-Mallory staining on paraffin-embedded sections of xenografts derived from the injection of CRC sphere cells and treated for 4 weeks (6–9 weeks) with PBS (vehicle) or BMP7v. Data are representative of three independent experiments using different CRC sphere cell lines (CSC#2, 7, and 18). b Percentage of necrosis evaluated on paraffin-embedded sections of xenografts treated as in a. Data are shown as mean ± SD of three independent experiments. c Immunohistochemical analysis of CD31 and VEGFR2 (red staining) on paraffin-embedded sections of xenografts generated by the injection of CRC sphere cell lines and treated with PBS (vehicle), BMP4, or BMP7v. Green arrowheads indicate microvessels expressing CD31 or VEGFR2. Images are representative of three independent experiments using cells as in a. Nuclei were revealed by hematoxylin staining (blue). The scale bar represents 20 µm. d Number of microvessels positive for CD31 (left panel) and VEGFR2 (right panel) expression, evaluated on paraffin-embedded sections of xenografts treated as in c. Data are shown as mean ± SD of cells. MVD = microvessel density. e Fold change of viable cells in 35 CR-CSC lines treated with oxaliplatin/5-FU for 24 h. Dotted line indicates the threshold between chemoresistant (red) and sensitive CR-CSCs (green). f Cell viability percentage in chemoresistant CR-CSCs (R1-R4) pretreated with BMP7 for 3 days and with oxaliplatin/5-FU (oxa/5-FU) for additional 24 h as indicated. Data are shown as mean ± SD of three different experiments performed in the indicated R-CSCs. g Colony forming efficiency of CR-CSCs treated as in f and evaluated at 21 days. Representative soft-agar analyses are reported in the lower part of the graph. Bars show the mean ± SD of seven different CRC sphere cell lines (CSC#1–3, 5, 7, 10, and 18). h Tumor size of subcutaneous growth of the indicated CR-CSCs. Mice were treated for 4 weeks (6–9 weeks) with vehicle, oxaliplatin/5-FU (oxa/5-FU) and BMP7v alone or in combination. Error bars show the mean ± SD of tumor size measured in six mice/group. Black arrowheads indicate days of treatment. i Immunohistochemical analysis of CD44v6, beta -catenin, Ki67, and CK20 (red color) in paraffin-embedded sections of CSC#7 xenografts treated as in h. Nuclei were counterstained by aqueous hematoxylin (blue color). The scale bar represents 20 µm (left panels). Percentage of CD44v6, beta -catenin, Ki67, and CK20 positive cells in paraffin-embedded sections of tumor xenografts treated with vehicle (V), BMP7v (B), oxaliplatin/5-FU (O/F), alone or in combination (B/O/F) for 72 h. Error bars are mean ± SD of positive cell counts in three serial embedded-paraffin sections of six tumor xenografts per group derived from the injection of three different CRC sphere cells (CSC#1, 2, and 7) (right panels) Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31591478), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human VEGFR2/KDR/Flk-1 by Western Blot

Detection of Human VEGFR2/KDR/Flk-1 by Western Blot

Exogenous VEGF165 suppresses RASSF1A expression in normal epithelial and endothelial cells. Metastatic colon cancer cells (mCRC, (A) T84 and (B) Colo 205) were stimulated with VEGF165 as indicated before and cell metabolism was performed by MTT assay for 6 h (black dash), 24 h (green) and 48 h (pink). Immortalized benign protate ((C) RWPE1), human bronchial epithelial cells ((D) HBEpc), immortalized human breast epithelial cells ((E) MCF-10A), endothelial cells (F, EC) were stimulated with or without VEGF165 (12.5 and 25 ng/mL) for 24 h. Cell lysates were monitored by Western blot for p-VEGFR1, p-VEGFR2, RASSF1A, and GAPDH as indicated. Immunoblot was quantified by scanning densitometry and normalized against GAPDH expression for RWPE1 (G), HBEpc (H), MCF 10A (I) and EC (lower panel of (F)). Results are from three independent experiments and statistical significance was determined using one way-ANOVA followed Bonferroni test. (* p < 0.05, ** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human VEGFR2/KDR/Flk-1 by Western Blot

Detection of Human VEGFR2/KDR/Flk-1 by Western Blot

Exogenous VEGF165 suppresses RASSF1A expression in normal epithelial and endothelial cells. Metastatic colon cancer cells (mCRC, (A) T84 and (B) Colo 205) were stimulated with VEGF165 as indicated before and cell metabolism was performed by MTT assay for 6 h (black dash), 24 h (green) and 48 h (pink). Immortalized benign protate ((C) RWPE1), human bronchial epithelial cells ((D) HBEpc), immortalized human breast epithelial cells ((E) MCF-10A), endothelial cells (F, EC) were stimulated with or without VEGF165 (12.5 and 25 ng/mL) for 24 h. Cell lysates were monitored by Western blot for p-VEGFR1, p-VEGFR2, RASSF1A, and GAPDH as indicated. Immunoblot was quantified by scanning densitometry and normalized against GAPDH expression for RWPE1 (G), HBEpc (H), MCF 10A (I) and EC (lower panel of (F)). Results are from three independent experiments and statistical significance was determined using one way-ANOVA followed Bonferroni test. (* p < 0.05, ** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human VEGFR2/KDR/Flk-1 by Western Blot

Detection of Human VEGFR2/KDR/Flk-1 by Western Blot

Exogenous VEGF165 suppresses RASSF1A expression in normal epithelial and endothelial cells. Metastatic colon cancer cells (mCRC, (A) T84 and (B) Colo 205) were stimulated with VEGF165 as indicated before and cell metabolism was performed by MTT assay for 6 h (black dash), 24 h (green) and 48 h (pink). Immortalized benign protate ((C) RWPE1), human bronchial epithelial cells ((D) HBEpc), immortalized human breast epithelial cells ((E) MCF-10A), endothelial cells (F, EC) were stimulated with or without VEGF165 (12.5 and 25 ng/mL) for 24 h. Cell lysates were monitored by Western blot for p-VEGFR1, p-VEGFR2, RASSF1A, and GAPDH as indicated. Immunoblot was quantified by scanning densitometry and normalized against GAPDH expression for RWPE1 (G), HBEpc (H), MCF 10A (I) and EC (lower panel of (F)). Results are from three independent experiments and statistical significance was determined using one way-ANOVA followed Bonferroni test. (* p < 0.05, ** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human VEGFR2/KDR/Flk-1 by Western Blot

Detection of Human VEGFR2/KDR/Flk-1 by Western Blot

VEGF-C induces LEC tube formation.A, Prostatic LEC tube length at 4.5 hours post VEGF-C treatment was quantified using ImageJ. VEGF-C significantly increased the number of tubes formed compared to vehicle control. Data expressed as mean±s.e.m., n = 3, ***P<0.001 using One-way ANOVA, Bonferroni post-analysis. B, Western blotting analysis of VEGFR-2 and VEGFR-3 expression in lung, neonatal dermis and prostate LECs. beta -tubulin was used as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/22745786), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human VEGFR2/KDR/Flk-1 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human placenta and kidney

Western Blot

1 µg/mL
Sample: HUVEC human umbilical vein endothelial cells

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.

Reviewed Applications

Read 6 reviews rated 3.8 using AF357 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

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

Long Name

Vascular Endothelial Growth Factor Receptor 2

Alternate Names

CD309, Flk-1, Flk1, KDR, KRD1, Ly73, VEGF R2

Entrez Gene IDs

3791 (Human); 16542 (Mouse)

Gene Symbol

KDR

UniProt

AAC16450

Additional VEGFR2/KDR/Flk-1 Products

Product Documents for Human VEGFR2/KDR/Flk-1 Antibody

Certificate of Analysis

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Product Specific Notices for Human VEGFR2/KDR/Flk-1 Antibody

For research use only

Citations for Human VEGFR2/KDR/Flk-1 Antibody

Customer Reviews for Human VEGFR2/KDR/Flk-1 Antibody (6)

3.8 out of 5
6 Customer Ratings
5 Stars
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Customer Images

Showing  1 - 5 of 6 reviews Showing All
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  • Human VEGFR2/KDR/Flk-1 Antibody
    Name: Karina Kinghorn
    Application: Western Blot
    Sample Tested: HUVEC human umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 08/04/2021
    First lane- nontargeting, second lane- AP1 knockdown. Top ladder band 220kDa.
    Human VEGFR2/KDR/Flk-1 Antibody AF357
  • Human VEGFR2/KDR/Flk-1 Antibody
    Name: Karina Kinghorn
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HUVEC human umbilical vein endothelial cells
    Species: Human
    Verified Customer | Posted 07/16/2021
    The staining pattern is much more punctate than the Cell Signaling VEGFR2 antibody with no VEGF added
    Human VEGFR2/KDR/Flk-1 Antibody AF357
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 19567589
    Species: Human
    Verified Customer | Posted 01/08/2015
  • Name: Anonymous
    Application: Immunoprecipitation
    Sample Tested: See PMID 19857463
    Species: Human
    Verified Customer | Posted 01/08/2015
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 23267826
    Species: Mouse
    Verified Customer | Posted 01/08/2015
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 23746980
    Species: Human
    Verified Customer | Posted 01/07/2015

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