Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

R&D Systems | Catalog # DYC2724-2

R&D Systems
100% Guarantee

Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Sample Type

Cell Lysates

Reactivity

Human

Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA Features

  • Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Available in 2, 5, and 15-(96-well) plate pack sizes
  • Economical alternative to Western blot
View all VEGFR3/Flt-4 ELISA Kits »

Product Summary for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated human VEGF R3 / Flt-4 in cell lysates. An immobilized capture antibody specific for VEGF R3 / Flt-4 binds both phosphorylated and unphosphorylated VEGF R3 / Flt-4. After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

Cell lysates (100 µL)

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

Figure 1. The Human Phospho-VEGF R3 DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western analysis

Lysates prepared from human VEGF R3 transfected NS0 cells (NS0-hVEGF R3) were diluted in a series and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were done using an anti-VEGF R3 monoclonal antibody and goat anti-mouse agarose. Immunoblots were incubated with a biotinylated anti-phosphotyrosine monoclonal antibody (Catalog # BAM1676) to detect phospho-VEGF R3. Bands were visualized with Streptavidin-HRP (Cat # DY998) followed by chemiluminescent detection using WesternGloTM Chemiluminescent Detection Substrate (Catalog # AR004). Human Phospho-VEGF R3 can be detected in this DuoSet IC ELISA by using approximately 10 to 15 times less lysate than is needed for a conventional IP-Western blot.

Figure 2. The specificity of the Human Phospho-VEGF R3 DuoSet IC ELISA is confirmed by receptor competition

The indicated amounts of recombinant extracellular domains of human VEGF R3 (Catalog #349-F4), human VEGF R1 (Catalog #321-FL), human VEGF R2 (Catalog #357-KD) or human Tie-2 (Catalog #313-TI) were added to 20 μg NS0-hVEGF R3 cell lysate and analyzed using this DuoSet IC ELISA. Competition was observed only with recombinant human VEGF R3.

Kit Contents for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Background: VEGFR3/Flt-4

VEGFR1 (Flt-1), VEGFR2 (KDR/Flk-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 domain and kinase insert domains in their intracellular region. They are best known for regulating VEGF family-mediated vasculogenesis, angiogenesis, and lymphangiogenesis. They are also mediators of neurotrophic activity and regulators of hematopoietic development. VEGFR2 is thought to be the primary inducer of VEGF-mediated blood vessel growth, while VEGFR3 plays a significant role in VEGF-C and VEGF-D-mediated lymphangiogenesis.

Long Name

Vascular Endothelial Growth Factor Receptor 3

Alternate Names

Flt-4, FLT4, VEGF R3

Entrez Gene IDs

2324 (Human); 14257 (Mouse)

Gene Symbol

FLT4

Additional VEGFR3/Flt-4 Products

Product Documents for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

For research use only

Citations for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

Customer Reviews for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA (2)

5 out of 5
2 Customer Ratings
5 Stars
100%
4 Stars
0%
3 Stars
0%
2 Stars
0%
1 Stars
0%

Have you used Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Customer Images

Showing  1 - 2 of 2 reviews Showing All
Filter By:
Clear Filters X
  • Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA
    Name: Anonymous
    Sample Tested: Breast cancer tissue and Serum and Plasma
    Verified Customer | Posted 10/29/2023
    Whether it's tissue specimens, blood samples, or supernatant, we can clearly see the differences and obtain reliable test results.
    Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA DYC2724-2
  • Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA
    Name: Jenna Nguyen
    Sample Tested: 7TD1 mouse hybridoma cell line
    Verified Customer | Posted 10/17/2019

There are no reviews that match your criteria.

Showing  1 - 2 of 2 reviews Showing All

FAQs for Human Phospho-VEGFR3/Flt-4 DuoSet IC ELISA

Showing  1 - 1 of 1 FAQ Showing All

  • Q: Which phosphorylated sites are recognized by this assay?

    A: This assay utilizes an anti-phosphorylated tyrosine monoclonal detection antibody, and it recognizes all phosphorylated tyrosine residues.

Showing  1 - 1 of 1 FAQ Showing All
View all FAQs for ELISA Kits