Human CDKN3 (Cyclin Dependent Kinase Inhibitor 3) CLIA Kit (HUES00233)
- Product type:
- ELISA Kit
- ELISA Type:
|Detection range:||31.25-2000 pg/mL|
|Sample type:||Tissue homogenates|
|Repeatability:||CV < 15%|
|Specificity:||This kit recognizes Human CDKN3 in samples. No significant cross-reactivity or interference between Human CDKN3 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 CDKN3. 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 CDKN3 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 CDKN3, 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 CDKN3. The concentration of Human CDKN3 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
|UniProt Protein Function:||CDKN3: May play a role in cell cycle regulation. Dual specificity phosphatase active toward substrates containing either phosphotyrosine or phosphoserine residues. Dephosphorylates CDK2 at 'Thr-160' in a cyclin-dependent manner. Defects in CDKN3 are found in patients with hepatocellular carcinoma (HCC). Belongs to the protein-tyrosine phosphatase family. 2 isoforms of the human protein are produced by alternative splicing.|
|UniProt Protein Details:|
Protein type:Motility/polarity/chemotaxis; Protein phosphatase, dual-specificity; EC 3. 1. 3. 16; Cell cycle regulation; EC 3. 1. 3. 48
Chromosomal Location of Human Ortholog: 14q22
Cellular Component: perinuclear region of cytoplasm; cytoplasm
Molecular Function:protein binding; protein tyrosine/serine/threonine phosphatase activity; protein tyrosine phosphatase activity; protein serine/threonine phosphatase activity
Biological Process: negative regulation of cell proliferation; regulation of cyclin-dependent protein kinase activity; cell cycle arrest; G1/S transition of mitotic cell cycle
Disease: Hepatocellular Carcinoma
|NCBI Summary:||The protein encoded by this gene belongs to the dual specificity protein phosphatase family. It was identified as a cyclin-dependent kinase inhibitor, and has been shown to interact with, and dephosphorylate CDK2 kinase, thus prevent the activation of CDK2 kinase. This gene was reported to be deleted, mutated, or overexpressed in several kinds of cancers. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Aug 2008]|
|NCBI GenInfo Identifier:||2499769|
|NCBI Gene ID:||1033|
|NCBI Accession:||Q16667. 1|
|UniProt Related Accession:||Q16667|
|Molecular Weight:||23805 Da|
|NCBI Full Name:||Cyclin-dependent kinase inhibitor 3|
|NCBI Synonym Full Names:||cyclin dependent kinase inhibitor 3|
|NCBI Official Symbol:||CDKN3|
|NCBI Official Synonym Symbols:||KAP; CDI1; CIP2; KAP1|
|NCBI Protein Information:||cyclin-dependent kinase inhibitor 3|
|UniProt Protein Name:||Cyclin-dependent kinase inhibitor 3|
|UniProt Synonym Protein Names:||CDK2-associated dual-specificity phosphatase; Cyclin-dependent kinase interactor 1; Cyclin-dependent kinase-interacting protein 2; Kinase-associated phosphatase|
|Protein Family:||Cyclin-dependent kinase inhibitor|
|UniProt Gene Name:||CDKN3|
|UniProt Entry Name:||CDKN3_HUMAN|
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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CDKN3 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CDKN3 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||8.96||10.33||10.47||11.19||7.01||10.12|
The recovery of Human CDKN3 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)||95-109||101|
|Cell culture media (n=5)||91-102||97|
Samples were spiked with high concentrations of Human CDKN3 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 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.