Human TGFBR2(TGF-beta receptor type 2) ELISA Kit (HUES03605)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||0.78-50 ng/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Human TGFBR2 in samples. No significant cross-reactivity or interference between Human TGFBR2 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 Human TGFBR2. 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 Human TGFBR2 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 TGFBR2, 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 Human TGFBR2. The concentration of Human TGFBR2 in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||TGFBR2: a TKL kinase of the serine/threonine-protein kinase receptor (STKR) family. R1 and R2 TGF-beta receptors dimerize after binding TGF-beta at the cell surface. Binds to DAXX. Defects can cause esophageal cancer.|
|UniProt Protein Details:|
Protein type:Oncoprotein; Membrane protein, integral; Protein kinase, TKL; EC 2. 7. 11. 30; Protein kinase, Ser/Thr (receptor); Kinase, protein; TKL group; STKR family; Type2 subfamily
Chromosomal Location of Human Ortholog: 3p22
Cellular Component: caveola; cytosol; external side of plasma membrane; integral to membrane; lipid raft; plasma membrane; receptor complex
Molecular Function:ATP binding; glycosaminoglycan binding; metal ion binding; mitogen-activated protein kinase kinase kinase binding; protein binding; receptor signaling protein serine/threonine kinase activity; SMAD binding; transforming growth factor beta binding; transforming growth factor beta receptor activity; transforming growth factor beta receptor activity, type II; transmembrane receptor protein serine/threonine kinase activity
Biological Process: activation of protein kinase activity; aging; apoptosis; blood vessel development; brain development; common-partner SMAD protein phosphorylation; embryo implantation; embryonic cranial skeleton morphogenesis; embryonic hemopoiesis; gastrulation; gut development; heart development; in utero embryonic development; lens development in camera-type eye; myeloid dendritic cell differentiation; negative regulation of cardiac muscle cell proliferation; negative regulation of transforming growth factor beta receptor signaling pathway; Notch signaling pathway; organ regeneration; palate development; patterning of blood vessels; peptidyl-serine phosphorylation; peptidyl-threonine phosphorylation; positive regulation of angiogenesis; positive regulation of B cell tolerance induction; positive regulation of cell proliferation; positive regulation of mesenchymal cell proliferation; positive regulation of NK T cell differentiation; positive regulation of skeletal muscle regeneration; positive regulation of smooth muscle cell proliferation; positive regulation of T cell tolerance induction; positive regulation of tolerance induction to self antigen; protein amino acid phosphorylation; receptor-mediated endocytosis; regulation of cell proliferation; regulation of gene expression; response to drug; response to estrogen stimulus; response to glucose stimulus; response to mechanical stimulus; response to nutrient; smoothened signaling pathway; transforming growth factor beta receptor signaling pathway; vasculogenesis; wound healing
Disease: Colorectal Cancer, Hereditary Nonpolyposis, Type 6; Esophageal Cancer; Loeys-dietz Syndrome 2
|NCBI Summary:||This gene encodes a member of the Ser/Thr protein kinase family and the TGFB receptor subfamily. The encoded protein is a transmembrane protein that has a protein kinase domain, forms a heterodimeric complex with another receptor protein, and binds TGF-beta. This receptor/ligand complex phosphorylates proteins, which then enter the nucleus and regulate the transcription of a subset of genes related to cell proliferation. Mutations in this gene have been associated with Marfan Syndrome, Loeys-Deitz Aortic Aneurysm Syndrome, and the development of various types of tumors. Alternatively spliced transcript variants encoding different isoforms have been characterized. [provided by RefSeq, Jul 2008]|
|NCBI GenInfo Identifier:||116242818|
|NCBI Gene ID:||7048|
|NCBI Accession:||P37173. 2|
|UniProt Secondary Accession:||P37173,Q15580, Q6DKT6, Q99474, B4DTV5,|
|UniProt Related Accession:||P37173|
|Molecular Weight:||67,457 Da|
|NCBI Full Name:||TGF-beta receptor type-2|
|NCBI Synonym Full Names:||transforming growth factor beta receptor II|
|NCBI Official Symbol:||TGFBR2|
|NCBI Official Synonym Symbols:||AAT3; FAA3; LDS2; MFS2; RIIC; LDS1B; LDS2B; TAAD2; TGFR-2; TGFbeta-RII|
|NCBI Protein Information:||TGF-beta receptor type-2|
|UniProt Protein Name:||TGF-beta receptor type-2|
|UniProt Synonym Protein Names:||TGF-beta type II receptor; Transforming growth factor-beta receptor type II; TGF-beta receptor type II; TbetaR-II|
|Protein Family:||TGF-beta receptor|
|UniProt Gene Name:||TGFBR2|
|UniProt Entry Name:||TGFR2_HUMAN|
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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TGFBR2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TGFBR2 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||6.47||4.88||4.86||5.60||4.11||4.91|
The recovery of Human TGFBR2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||86-102||94|
|Cell culture media (n=5)||86-100||92|
Samples were spiked with high concentrations of Human TGFBR2 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)|
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.
|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.