|Detection Range:||0.31-20 ng/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Human PTK6 in samples. No significant cross-reactivity or interference between Human PTK6 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 PTK6. 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 PTK6 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 PTK6, 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 PTK6. The concentration of Human PTK6 in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||Brk: a tyrosine kinase of the Src family. May function as an intracellular signal transducer in epithelial tissues. Very high level expression in colon and high levels in small intestine and prostate, and low levels in some fetal tissues. Selectively expressed in breast tumors and cell lines, and perhaps in colon and prostate cancers. Also found in melanocytes. Not expressed in heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Overexpression in mammary cells leads to EGF sensitization and results in a partially transformed phenotype. Enhances anchorage-independent growth and responsiveness to EGF. RNAi reduces proliferation in breast cancer cells. Kinase-inactive mutant indicates tumor function may be independent of catalytic function.|
|UniProt Protein Details:|
Protein type:EC 2. 7. 10. 2; Kinase, protein; Protein kinase, TK; Protein kinase, tyrosine (non-receptor); TK group; Src family
Chromosomal Location of Human Ortholog: 20q13. 3
Cellular Component: nucleoplasm; ruffle; extrinsic to internal side of plasma membrane; cytoplasm; nucleus
Molecular Function:identical protein binding; protein binding; non-membrane spanning protein tyrosine kinase activity; ATP binding; receptor binding
Biological Process: cell migration; protein amino acid autophosphorylation; tyrosine phosphorylation of Stat3 protein; innate immune response; actin cytoskeleton organization and biogenesis; tyrosine phosphorylation of Stat5 protein; protein amino acid phosphorylation; transmembrane receptor protein tyrosine kinase signaling pathway; negative regulation of growth; regulation of cell proliferation
|NCBI Summary:||The protein encoded by this gene is a cytoplasmic nonreceptor protein kinase which may function as an intracellular signal transducer in epithelial tissues. Overexpression of this gene in mammary epithelial cells leads to sensitization of the cells to epidermal growth factor and results in a partially transformed phenotype. Expression of this gene has been detected at low levels in some breast tumors but not in normal breast tissue. The encoded protein has been shown to undergo autophosphorylation. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Jan 2012]|
|NCBI GenInfo Identifier:||8928302|
|NCBI Gene ID:||5753|
|NCBI Accession:||Q13882. 1|
|UniProt Secondary Accession:||Q13882,Q58F01, B2RCR3, B4DW46,|
|UniProt Related Accession:||Q13882|
|NCBI Full Name:||Protein-tyrosine kinase 6|
|NCBI Synonym Full Names:||protein tyrosine kinase 6|
|NCBI Official Symbol:||PTK6|
|NCBI Official Synonym Symbols:||BRK|
|NCBI Protein Information:||protein-tyrosine kinase 6; breast tumor kinase; protein-tyrosine kinase BRK; tyrosine-protein kinase BRK; PTK6 protein tyrosine kinase 6|
|UniProt Protein Name:||Protein-tyrosine kinase 6|
|UniProt Synonym Protein Names:||Breast tumor kinase; Tyrosine-protein kinase BRK|
|Protein Family:||Protein-tyrosine kinase|
|UniProt Gene Name:||PTK6|
|UniProt Entry Name:||PTK6_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 PTK6 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PTK6 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.00||4.51||4.92||6.48||5.13||4.11|
The recovery of Human PTK6 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)||85-98||92|
|Cell culture media (n=5)||86-99||92|
Samples were spiked with high concentrations of Human PTK6 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.