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Human Total VEGFR2/KDR

Human Total VEGFR2/KDR DuoSet IC ELISA Summary

Assay   Type

Solid   Phase Sandwich ELISA

Format

96-well   strip plate

Assay   Length

4 hours   40 minutes (after plate preparation)

Sample   Type & Volume Required Per Well

Cell   lysates (100 μL)

Sensitivity

0.018   ng/mL

Assay   Range

62.5 -   4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)

Specificity

Total   VEGF R2 / KDR / Flk-1 in cell lysates

Cross-reactivity

<   0.5% cross-reactivity observed with available related molecules.< 50%   cross-species reactivity observed with species tested

Interference

No   significant interference observed with available related molecules.

Product Summary

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total VEGF R2 / KDR / Flk-1 in cell lysates. An immobilized capture antibody specific for VEGF R2 / KDR / Flk-1 binds both phosphorylated and unphosphorylated VEGF R2 / KDR / Flk-1. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.

Preparation and Storage

Shipping

The   product is shipped at ambient temperature. Upon receipt, store it immediately   at the temperature recommended below.

Storage

Store   the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: VEGFR2/KDR/Flk-1

VEGF R1 (Flt-1), VEGF R2 (KDR/Flk-1), and VEGF R3 (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. VEGF R2 is thought to be the primary inducer of VEGF-mediated blood vessel growth, while VEGF R3 plays a significant role in VEGF-C and VEGF-D-mediated lymphangiogenesis.

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

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3Add 50 μL of Assay Diluent to each well.

4Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7Aspirate and wash 5 times.

8Add 100 μL Substrate Solution to each well.

9Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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