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Human VEGFR1/Flt-1

Human VEGFR1/Flt-1 Quantikine ELISA Kit Summary

Assay   Type

Solid   Phase Sandwich ELISA

Format

96-well   strip plate

Assay   Length

4.5   hours

Sample   Type & Volume Required Per Well

Cell   Culture Supernates (100 uL), Cell Lysates (10 uL), Serum (100 uL), EDTA   Plasma (100 uL), Heparin Plasma (100 uL), Saliva (100 uL)

Sensitivity

8.46   pg/mL

Assay   Range

31.3 -   2,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma,   Heparin Plasma, Saliva)

Specificity

Natural   and recombinant human VEGF R1

 

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

The Quantikine® Human VEGF R1/Flt-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human VEGF R1 in cell culture supernates, cell lysates, serum, plasma, and saliva. It contains NS0-expressed recombinant human VEGF R1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human VEGF R1 showed linear curves that were parallel to the standard curves obtained using the Quantikine® kit standards. These results indicate that this kit can be used to determine relative mass values for natural human VEGF R1.

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: VEGFR1/Flt-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 1

Entrez   Gene IDs:

2321   (Human); 14254 (Mouse)

Alternate   Names:

EC   2.7.10; EC 2.7.10.1; FLT; FLT1; Flt-1; Fms-like tyrosine kinase 1;   fms-related tyrosine kinase 1 (vascular endothelial growth   factor/vascularpermeability factor receptor); FRT; Tyrosine-protein kinase   FRT; Tyrosine-protein kinase receptor FLT; vascular endothelial growth factor   receptor 1; Vascular permeability factor receptor; VEGF R1; VEGFR1; VEGFR-1

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.

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

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

3.     Add 50 μL of Assay Diluent to each well.

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

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

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

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 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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