Rat Immunology ELISA Kits 3
Rat VEGFR2 / VEGF Receptor 2 ELISA Kit
- SKU:
- RTFI01209
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- O08775
- Sensitivity:
- 0.094ng/ml
- Range:
- 0.156-10ng/ml
- ELISA Type:
- Sandwich
- Synonyms:
- VEGFR2, KDR, VEGFR, VEGFR-2, CD309, Flk-1, CD309 antigen, Fetal liver kinase 1, fetal liver kinase-1, FLK-1, FLK1tyrosine kinase growth factor receptor, Kinase insert domain receptor, kinase insert domain receptor, a type III receptor tyrosine kinase
- Reactivity:
- Rat
- Research Area:
- Cardiovascular
Description
| Product Name: | Rat VEGFR-2/KDR (Vascular Endothelial Growth Factor Receptor 2) ELISA Kit |
| Product Code: | RTFI01209 |
| Size: | 96 Assays |
| Target: | Rat VEGFR-2/KDR |
| Alias: | VEGFR2, KDR, VEGFR, VEGFR-2, CD309, Flk-1, CD309 antigen, Fetal liver kinase 1, fetal liver kinase-1, FLK-1, FLK1tyrosine kinase growth factor receptor, Kinase insert domain receptor, kinase insert domain receptor, a type III receptor tyrosine kinase, Protein-tyrosine kinase receptor flk-1, soluble VEGFR2, vascular endothelial growth factor receptor 2 |
| Reactivity: | Rat |
| Detection Method: | Sandwich ELISA, Double Antibody |
| Sensitivity: | 0.094ng/ml |
| Range: | 0.156-10ng/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
| Recovery: | Matrices listed below were spiked with certain level of Rat VEGFR-2/KDR and the recovery rates were calculated by comparing the measured value to the expected amount of Rat VEGFR-2/KDR in samples. | ||||||||||||||||
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| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat VEGFR-2/KDR and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. | ||||||||||||||||
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| Intra-Assay: | CV <8% | ||||||||||||||||
| Inter-Assay: | CV <10% |
| Uniprot: | O08775 |
| 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:EC 2.7.10.1; Protein kinase, tyrosine (receptor); Kinase, protein; Protein kinase, TK; Membrane protein, integral; TK group; VEGFR family Cellular Component: Golgi apparatus; neuron projection; cell surface; integral to plasma membrane; endoplasmic reticulum; early endosome; cytoplasmic membrane-bound vesicle; lipid raft; cell soma; cytoplasm; plasma membrane; nucleus; cell junction; endosome; external side of plasma membrane Molecular Function:integrin binding; vascular endothelial growth factor receptor activity; protein binding; protein-tyrosine kinase activity; growth factor binding; ATP binding Biological Process: positive regulation of positive chemotaxis; peptidyl-tyrosine phosphorylation; protein amino acid autophosphorylation; cell maturation; positive regulation of calcium-mediated signaling; positive regulation of vasodilation; positive regulation of long-term neuronal synaptic plasticity; calcium ion homeostasis; regulation of cell shape; elevation of cytosolic calcium ion concentration; positive regulation of MAPKKK cascade; positive regulation of focal adhesion formation; ovarian follicle development; positive regulation of cell proliferation; positive regulation of mesenchymal cell proliferation; hemopoiesis; angiogenesis; negative regulation of neuron apoptosis; vasculogenesis; positive regulation of TOR signaling pathway; positive regulation of BMP signaling pathway; aging; response to drug; endothelial cell differentiation; cell migration; positive regulation of neurogenesis; cell fate commitment; embryonic hemopoiesis; male gonad development; regulation of endothelial cell differentiation; positive regulation of phosphoinositide 3-kinase cascade; positive regulation of angiogenesis; branching morphogenesis of a tube; cell migration during sprouting angiogenesis; response to hypoxia; positive regulation of endothelial cell proliferation; positive regulation of protein amino acid phosphorylation; vascular endothelial growth factor receptor signaling pathway; lymph vessel development; surfactant homeostasis; hyaluronan metabolic process; alveolus development; neurite morphogenesis; positive regulation of epithelial cell proliferation; positive regulation of cell migration; negative regulation of apoptosis; lung development; positive regulation of nitric-oxide synthase biosynthetic process |
| NCBI Summary: | receptor for vascular endothelial growth factor (VEGF) [RGD, Feb 2006] |
| UniProt Code: | O08775 |
| NCBI GenInfo Identifier: | 6981128 |
| NCBI Gene ID: | 25589 |
| NCBI Accession: | NP_037194.1 |
| UniProt Related Accession: | O08775 |
| Molecular Weight: | 150,394 Da |
| NCBI Full Name: | vascular endothelial growth factor receptor 2 |
| NCBI Synonym Full Names: | kinase insert domain receptor |
| NCBI Official Symbol: | Kdr |
| NCBI Official Synonym Symbols: | Vegfr-2 |
| NCBI Protein Information: | vascular endothelial growth factor receptor 2; FLK-1; fetal liver kinase 1; kinase insert domain protein receptor; protein-tyrosine kinase receptor flk-1; FLK1 kinase insert domain receptor (VEGF receptor 2); FLK1 kinase insert domain receptor (a type III receptor tyrosine kinase) (VEGF receptor 2) |
| UniProt Protein Name: | Vascular endothelial growth factor receptor 2 |
| UniProt Synonym Protein Names: | Fetal liver kinase 1; FLK-1; Protein-tyrosine kinase receptor flk-1 |
| Protein Family: | Vascular endothelial growth factor receptor |
| UniProt Gene Name: | Kdr |
| UniProt Entry Name: | VGFR2_RAT |
| Step | Procedure |
| 1. | Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
| 2. | Aliquot 0.1ml standard solutions into the standard wells. |
| 3. | Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. |
| 4. | Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. |
| 5. | Seal the plate with a cover and incubate at 37°C for 90 min. |
| 6. | Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. |
| 7. | Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. |
| 8. | Seal the plate with a cover and incubate at 37°C for 60 min. |
| 9. | Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
| 10. | Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min. |
| 11. | Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. |
| 12. | Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. |
| 13. | Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
| 14. | Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum: | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clotovernight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Removeserum and assay promptly or aliquot and store the samples at-80°C. Avoid multiple freeze-thaw cycles. |
| Plasma: | Collect plasma using EDTA or heparin as an anti-coagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.Note: Over haemolysed samples are not suitable for use with this kit. |
| Urine & Cerebrospinal Fluid: | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
| Cell Culture Supernatant: | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
| Cell Lysates: | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20°C. |
| Tissue Homogenates: | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenizein 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or-80°C. |
| Tissue Lysates: | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Breast Milk: | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
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