Mouse VEGFR-2/KDR (Vascuoar Endothelial Growth Factor Receptor 2) ELISA Kit (MOES01156)
- SKU:
- MOES01156
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P35918
- Sensitivity:
- 0.09ng/mL
- Range:
- 0.16-10ng/mL
- ELISA Type:
- Sandwich
- Synonyms:
- VEGFR2, CD309, FLK1
- Reactivity:
- Mouse
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cardiovascular
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Mouse |
| Detection Method: | Colormetric |
| Detection Range: | 0.16-10 ng/mL |
| Sensitivity: | 0.10 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Mouse VEGFR-2/KDR in samples. No significant cross-reactivity or interference between Mouse VEGFR-2/KDR and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse VEGFR-2/KDR. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse VEGFR-2/KDR and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse VEGFR-2/KDR, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse VEGFR-2/KDR. The concentration of Mouse VEGFR-2/KDR in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | VEGFR2: a receptor tyrosine kinase of the VEGFR family. High affinity receptor for VEGF and VEGF-C. Ligand binding induces autophosphorylation and activation. Activated receptor recruits proteins including Shc, GRB2, PI3K, Nck, SHP-1 and SHP-2. Plays a key role in vascular development and regulation of vascular permeability. |
| UniProt Protein Details: | Protein type:Protein kinase, tyrosine (receptor); Kinase, protein; EC 2. 7. 10. 1; Protein kinase, TK; Membrane protein, integral; TK group; VEGFR family Cellular Component: Golgi apparatus; neuron projection; cell surface; endoplasmic reticulum; integral to plasma membrane; early endosome; extracellular region; integral to membrane; cytosol; lipid raft; membrane; cell soma; cytoplasm; plasma membrane; cytoplasmic vesicle; nucleus; cell junction; endosome; external side of plasma membrane Molecular Function:integrin binding; transferase activity; vascular endothelial growth factor receptor activity; protein binding; protein-tyrosine kinase activity; growth factor binding; transferase activity, transferring phosphorus-containing groups; nucleotide binding; kinase activity; transmembrane receptor protein tyrosine kinase activity; ATP binding; protein kinase activity Biological Process: positive regulation of positive chemotaxis; multicellular organismal development; cell maturation; positive regulation of vasodilation; positive regulation of long-term neuronal synaptic plasticity; protein amino acid phosphorylation; calcium ion homeostasis; regulation of cell shape; elevation of cytosolic calcium ion concentration; positive regulation of mesenchymal cell proliferation; negative regulation of neuron apoptosis; positive regulation of TOR signaling pathway; response to drug; regulation of endothelial cell proliferation; cell fate commitment; embryonic hemopoiesis; regulation of endothelial cell differentiation; positive regulation of phosphoinositide 3-kinase cascade; positive regulation of angiogenesis; cell migration during sprouting angiogenesis; branching morphogenesis of a tube; positive regulation of endothelial cell proliferation; lymph vessel development; surfactant homeostasis; alveolus development; transmembrane receptor protein tyrosine kinase signaling pathway; positive regulation of epithelial cell proliferation; positive regulation of nitric-oxide synthase biosynthetic process; negative regulation of apoptosis; peptidyl-tyrosine phosphorylation; protein amino acid autophosphorylation; positive regulation of calcium-mediated signaling; positive regulation of vascular endothelial growth factor receptor signaling pathway; positive regulation of MAPKKK cascade; positive regulation of focal adhesion formation; ovarian follicle development; positive regulation of cell proliferation; hemopoiesis; angiogenesis; vasculogenesis; cell differentiation; positive regulation of BMP signaling pathway; endothelial cell differentiation; cell migration; male gonad development; response to hypoxia; positive regulation of protein amino acid phosphorylation; vascular endothelial growth factor receptor signaling pathway; phosphorylation; neurite morphogenesis; positive regulation of cell migration; lung development |
| UniProt Code: | P35918 |
| NCBI GenInfo Identifier: | 549321 |
| NCBI Gene ID: | 16542 |
| NCBI Accession: | P35918. 1 |
| UniProt Secondary Accession: | P35918,C5H7S5, |
| UniProt Related Accession: | P35918 |
| Molecular Weight: | 75,230 Da |
| NCBI Full Name: | Vascular endothelial growth factor receptor 2 |
| NCBI Synonym Full Names: | kinase insert domain protein receptor |
| NCBI Official Symbol: | Kdr |
| NCBI Official Synonym Symbols: | Flk1; Ly73; Flk-1; Krd-1; VEGFR2; VEGFR-2; sVEGFR-2; 6130401C07 |
| NCBI Protein Information: | vascular endothelial growth factor receptor 2; kinase NYK; VEGF receptor-2; fetal liver kinase 1; protein-tyrosine kinase receptor flk-1; vascular endothelial growth factor receptor-2; vascular endothelial growth factor receptor-3; vascular endothelial growth factor receptor- 2; soluble vascular endothelial growth factor receptor 2 |
| UniProt Protein Name: | Vascular endothelial growth factor receptor 2 |
| UniProt Synonym Protein Names: | Fetal liver kinase 1; FLK-1; Kinase NYK; Protein-tyrosine kinase receptor flk-1; CD_antigen: CD309 |
| Protein Family: | Vascular endothelial growth factor receptor |
| UniProt Gene Name: | Kdr |
| UniProt Entry Name: | VGFR2_MOUSE |
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (ng/mL) | O.D | Average | Corrected |
| 10 | 2.455 2.483 | 2.469 | 2.398 |
| 5 | 1.563 1.589 | 1.576 | 1.505 |
| 2.5 | 0.965 0.963 | 0.964 | 0.893 |
| 1.25 | 0.463 0.491 | 0.477 | 0.406 |
| 0.63 | 0.247 0.235 | 0.241 | 0.17 |
| 0.31 | 0.186 0.156 | 0.171 | 0.1 |
| 0.16 | 0.123 0.125 | 0.124 | 0.053 |
| 0 | 0.066 0.076 | 0.071 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse VEGFR-2/KDR were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse VEGFR-2/KDR were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 0.50 | 1.30 | 4.00 | 0.50 | 1.40 | 3.60 |
| Standard deviation | 0.03 | 0.06 | 0.13 | 0.03 | 0.08 | 0.12 |
| C V (%) | 6.00 | 4.62 | 3.25 | 6.00 | 5.71 | 3.33 |
Recovery
The recovery of Mouse VEGFR-2/KDR spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 93-105 | 99 |
| EDTA plasma (n=5) | 85-98 | 92 |
| Cell culture media (n=5) | 94-110 | 101 |
Linearity
Samples were spiked with high concentrations of Mouse VEGFR-2/KDR and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 96-109 | 89-103 | 98-110 |
| Average (%) | 101 | 96 | 104 | |
| 1:4 | Range (%) | 95-109 | 83-93 | 98-111 |
| Average (%) | 102 | 88 | 104 | |
| 1:8 | Range (%) | 100-117 | 83-93 | 93-106 |
| Average (%) | 108 | 88 | 99 | |
| 1:16 | Range (%) | 98-112 | 87-101 | 94-107 |
| Average (%) | 104 | 94 | 100 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro ELISA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
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