Human Cell Biology ELISA Kits 4
Human PDGFR beta (Platelet Derived Growth Factor Receptor Beta) CLIA Kit (HUES00582)
- SKU:
- HUES00582
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 18.75pg/mL
- Range:
- 31.25-2000pg/mL
- ELISA Type:
- Sandwich
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 31.25-2000 pg/mL |
| Sensitivity: | 18.75 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human PDGFR beta in samples. No significant cross-reactivity or interference between Human PDGFR beta and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human PDGFR beta. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human PDGFR beta 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 Human PDGFR beta, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human PDGFR beta. The concentration of Human PDGFR beta in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | PDGFRB: a receptor tyrosine kinase of the PDGFR family that binds members of the platelet-derived growth factor family. The identity of the growth factor bound determines whether the functional receptor is a homodimer or a heterodimer, composed of both PDGFR-alpha and -beta. Ligand binding induces receptor dimerization and autophosphorylation, thereby recruiting SH2-containing proteins including Grb2, Src, GAP, PTPN11, PI3 kinase, PLC-gamma and Nck. Regulates cell growth, actin reorganization, migration and differentiation. A variety of myeloproliferative disorders and cancers result from translocations that activate PDGFRbeta by fusion with proteins such as TEL/ETV6, H2, CEV14/TRP11, rabaptin 5, and huntington interacting protein 1. Gleevec treatment of TEL fusions has been successful. Overexpressed in metastatic medulloblastoma. Inhibitors: Gleevec, Sutent. |
| UniProt Protein Details: | Protein type:EC 2. 7. 10. 1; Kinase, protein; Membrane protein, integral; Oncoprotein; PDGFR family; Protein kinase, TK; Protein kinase, tyrosine (receptor); TK group Chromosomal Location of Human Ortholog: 5q32 Cellular Component: apical plasma membrane; cytoplasm; focal adhesion; intrinsic to plasma membrane; membrane; nucleus; plasma membrane Molecular Function:enzyme binding; phosphatidylinositol-4,5-bisphosphate 3-kinase activity; platelet activating factor receptor activity; platelet-derived growth factor beta-receptor activity; platelet-derived growth factor binding; platelet-derived growth factor receptor activity; platelet-derived growth factor receptor binding; protein binding; protein kinase binding; protein-tyrosine kinase activity; Ras guanyl-nucleotide exchange factor activity; receptor binding Biological Process: cardiac myofibril assembly; cell migration; MAPKKK cascade; peptidyl-tyrosine phosphorylation; phosphatidylinositol metabolic process; phosphoinositide-mediated signaling; platelet-derived growth factor receptor signaling pathway; positive regulation of cell migration; positive regulation of cell proliferation; positive regulation of chemotaxis; positive regulation of MAP kinase activity; positive regulation of mitosis; positive regulation of phosphoinositide 3-kinase activity; positive regulation of phosphoinositide 3-kinase cascade; positive regulation of phosphoprotein phosphatase activity; positive regulation of smooth muscle cell migration; positive regulation of smooth muscle cell proliferation; protein amino acid autophosphorylation; regulation of actin cytoskeleton organization and biogenesis; regulation of phosphoinositide 3-kinase cascade; signal transduction Disease: Basal Ganglia Calcification, Idiopathic, 1; Basal Ganglia Calcification, Idiopathic, 4; Kosaki Overgrowth Syndrome; Myeloproliferative Disorder, Chronic, With Eosinophilia; Myofibromatosis, Infantile, 1; Premature Aging Syndrome, Penttinen Type |
| NCBI Summary: | This gene encodes a cell surface tyrosine kinase receptor for members of the platelet-derived growth factor family. These growth factors are mitogens for cells of mesenchymal origin. The identity of the growth factor bound to a receptor monomer determines whether the functional receptor is a homodimer or a heterodimer, composed of both platelet-derived growth factor receptor alpha and beta polypeptides. This gene is flanked on chromosome 5 by the genes for granulocyte-macrophage colony-stimulating factor and macrophage-colony stimulating factor receptor; all three genes may be implicated in the 5-q syndrome. A translocation between chromosomes 5 and 12, that fuses this gene to that of the translocation, ETV6, leukemia gene, results in chronic myeloproliferative disorder with eosinophilia. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P09619 |
| NCBI GenInfo Identifier: | 129890 |
| NCBI Gene ID: | 5159 |
| NCBI Accession: | P09619. 1 |
| UniProt Secondary Accession: | P09619,Q8N5L4, B5A957, |
| UniProt Related Accession: | P09619 |
| Molecular Weight: | Calculated: 37kDa/123kDaObserved: 170kDa |
| NCBI Full Name: | Platelet-derived growth factor receptor beta |
| NCBI Synonym Full Names: | platelet derived growth factor receptor beta |
| NCBI Official Symbol: | PDGFRB |
| NCBI Official Synonym Symbols: | IMF1; KOGS; IBGC4; JTK12; PDGFR; PENTT; CD140B; PDGFR1; PDGFR-1 |
| NCBI Protein Information: | platelet-derived growth factor receptor beta |
| UniProt Protein Name: | Platelet-derived growth factor receptor beta |
| UniProt Synonym Protein Names: | Beta platelet-derived growth factor receptor; Beta-type platelet-derived growth factor receptor; CD140 antigen-like family member B; Platelet-derived growth factor receptor 1; PDGFR-1; CD_antigen: CD140b |
| Protein Family: | Platelet-derived growth factor receptor |
| UniProt Gene Name: | PDGFRB |
As the RLU 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 (pg/mL) | RLU | Average | Corrected |
| 2000 | 52399 59043 | 55721 | 55693 |
| 1000 | 21854 26094 | 23974 | 23946 |
| 500 | 12011 10053 | 11032 | 11004 |
| 250 | 5164 5424 | 5294 | 5266 |
| 125 | 2747 2469 | 2608 | 2580 |
| 62.5 | 1387 1235 | 1311 | 1283 |
| 31.25 | 629 719 | 674 | 646 |
| 0 | 27 29 | 28 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PDGFR beta were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PDGFR beta 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 (pg/mL) | 106.49 | 212.77 | 704.78 | 97.43 | 215.77 | 727.00 |
| Standard deviation | 10.92 | 15.30 | 61.25 | 11.23 | 21.38 | 68.19 |
| C V (%) | 10.25 | 7.19 | 8.69 | 11.53 | 9.91 | 9.38 |
Recovery
The recovery of Human PDGFR beta 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) | 90-103 | 97 |
| EDTA plasma (n=5) | 89-101 | 94 |
| Cell culture media (n=5) | 86-99 | 91 |
Linearity
Samples were spiked with high concentrations of Human PDGFR beta 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 (%) | 85-96 | 89-100 | 91-102 |
| Average (%) | 91 | 95 | 97 | |
| 1:4 | Range (%) | 99-113 | 91-102 | 89-100 |
| Average (%) | 107 | 96 | 95 | |
| 1:8 | Range (%) | 95-111 | 101-118 | 98-113 |
| Average (%) | 102 | 108 | 105 | |
| 1:16 | Range (%) | 98-113 | 85-100 | 94-110 |
| Average (%) | 105 | 91 | 102 |
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 CLIA 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 A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| 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 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.
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