Description
Human CDK1 (Cyclin Dependent Kinase 1) ELISA Kit
The Human CDK1 (Cyclin-Dependent Kinase 1) ELISA Kit is a highly sensitive and specific assay designed for the quantitative measurement of CDK1 levels in human samples such as serum, plasma, and cell culture supernatants. This kit provides accurate and reproducible results, making it a valuable tool for a wide range of research applications.CDK1 is a key regulator of cell division and plays a critical role in the cell cycle progression. Dysregulation of CDK1 activity has been implicated in various diseases, including cancer, making it a promising target for therapeutics and biomarker development.
By using the Human CDK1 ELISA Kit, researchers can gain valuable insights into the role of CDK1 in disease pathogenesis and therapeutic response. This kit offers high-performance and reliability, making it an essential tool for studying CDK1 biology and its implications in human health and disease.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 0.16-10 ng/mL |
| Sensitivity: | 0.10 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human CDK1 in samples. No significant cross-reactivity or interference between Human CDK1 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 CDK1. 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 CDK1 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 CDK1, 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 CDK1. The concentration of Human CDK1 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | CDK1: a protein kinase of the CDK family. Catalytic subunit of the conserved protein complex known as M-phase promoting factor (MPF), which is essential for G1/S and G2/M phase transitions. Mitotic cyclins stably associate with this protein and function as regulatory subunits. Its activity is controlled by cyclin availability and phosphorylation through the cell cycle. Activated in many cancers including colon, liver and breast. The T isoform, which lacks a regulatory region, is expressed in breast cancer. Inhibition in cancer cells may drive cells into apoptosis. May also drive cell migration. Inhibitors: BMS-265246, BMS-265246-01. 2 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Cell cycle regulation; EC 2. 7. 11. 23; EC 2. 7. 11. 22; Motility/polarity/chemotaxis; Kinase, protein; Protein kinase, CMGC; Protein kinase, Ser/Thr (non-receptor); CMGC group; CDK family; CDK/CDK1 subfamily; CDK1 subfamily Chromosomal Location of Human Ortholog: 10q21. 1 Cellular Component: nucleoplasm; centrosome; membrane; mitochondrion; spindle microtubule; cytoplasm; midbody; nucleus; cytosol Molecular Function:RNA polymerase subunit kinase activity; protein serine/threonine kinase activity; protein binding; cyclin binding; cyclin-dependent protein kinase activity; histone kinase activity; Hsp70 protein binding; ATP binding; protein kinase activity Biological Process: regulation of Schwann cell differentiation; activation of MAPKK activity; nerve growth factor receptor signaling pathway; activation of MAPK activity; histone phosphorylation; response to toxin; ventricular cardiac muscle cell development; regulation of embryonic development; centrosome cycle; stress-activated MAPK cascade; toll-like receptor 3 signaling pathway; response to organic cyclic substance; toll-like receptor 5 signaling pathway; small GTPase mediated signal transduction; protein complex assembly; G2/M transition of mitotic cell cycle; toll-like receptor 4 signaling pathway; positive regulation of cardiac muscle cell proliferation; response to drug; mitosis; organ regeneration; fibroblast growth factor receptor signaling pathway; positive regulation of protein import into nucleus, translocation; cell aging; pronuclear fusion; toll-like receptor 2 signaling pathway; chromosome condensation; response to ethanol; cell division; response to activity; toll-like receptor 9 signaling pathway; response to amine stimulus; G1/S transition of mitotic cell cycle; negative regulation of apoptosis; positive regulation of ubiquitin-protein ligase activity during mitotic cell cycle; axon guidance; apoptosis; mitotic nuclear envelope disassembly; positive regulation of mitotic cell cycle; toll-like receptor 10 signaling pathway; epithelial cell differentiation; anaphase-promoting complex-dependent proteasomal ubiquitin-dependent protein catabolic process; response to axon injury; DNA replication; epidermal growth factor receptor signaling pathway; cell migration; MyD88-independent toll-like receptor signaling pathway; organelle organization and biogenesis; MAPKKK cascade; peptidyl-threonine phosphorylation; microtubule cytoskeleton organization and biogenesis; DNA repair; MyD88-dependent toll-like receptor signaling pathway; G1/S-specific transcription in mitotic cell cycle; peptidyl-serine phosphorylation; response to cadmium ion; regulation of ubiquitin-protein ligase activity during mitotic cell cycle; response to copper ion; mitotic cell cycle G2/M transition DNA damage checkpoint; Ras protein signal transduction; insulin receptor signaling pathway; toll-like receptor signaling pathway; innate immune response; mitotic cell cycle; vascular endothelial growth factor receptor signaling pathway; positive regulation of DNA replication |
| NCBI Summary: | The protein encoded by this gene is a member of the Ser/Thr protein kinase family. This protein is a catalytic subunit of the highly conserved protein kinase complex known as M-phase promoting factor (MPF), which is essential for G1/S and G2/M phase transitions of eukaryotic cell cycle. Mitotic cyclins stably associate with this protein and function as regulatory subunits. The kinase activity of this protein is controlled by cyclin accumulation and destruction through the cell cycle. The phosphorylation and dephosphorylation of this protein also play important regulatory roles in cell cycle control. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Mar 2009] |
| UniProt Code: | P06493 |
| NCBI GenInfo Identifier: | 334302921 |
| NCBI Gene ID: | 983 |
| NCBI Accession: | P06493. 3 |
| UniProt Secondary Accession: | P06493,O60764, A8K7C4, C9J497, |
| UniProt Related Accession: | P06493 |
| Molecular Weight: | 297 |
| NCBI Full Name: | Cyclin-dependent kinase 1 |
| NCBI Synonym Full Names: | cyclin-dependent kinase 1 |
| NCBI Official Symbol: | CDK1 |
| NCBI Official Synonym Symbols: | CDC2; CDC28A; P34CDC2 |
| NCBI Protein Information: | cyclin-dependent kinase 1; p34 protein kinase; cell cycle controller CDC2; cell division protein kinase 1; cell division control protein 2 homolog; cell division cycle 2, G1 to S and G2 to M |
| UniProt Protein Name: | Cyclin-dependent kinase 1 |
| UniProt Synonym Protein Names: | Cell division control protein 2 homolog; Cell division protein kinase 1; p34 protein kinase |
| UniProt Gene Name: | CDK1 |
| UniProt Entry Name: | CDK1_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.
| Concentration (ng/mL) | O.D | Average | Corrected |
| 10 | 2.421 2.425 | 2.423 | 2.358 |
| 5 | 1.702 1.742 | 1.722 | 1.657 |
| 2.5 | 1.001 0.987 | 0.994 | 0.929 |
| 1.25 | 0.504 0.538 | 0.521 | 0.456 |
| 0.63 | 0.272 0.264 | 0.268 | 0.203 |
| 0.32 | 0.183 0.161 | 0.172 | 0.107 |
| 0.16 | 0.111 0.127 | 0.119 | 0.054 |
| 0 | 0.065 0.065 | 0.065 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CDK1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CDK1 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 (ng/mL) | 0.50 | 0.86 | 4.72 | 0.46 | 0.91 | 4.66 |
| Standard deviation | 0.03 | 0.04 | 0.24 | 0.03 | 0.05 | 0.25 |
| C V (%) | 6.00 | 4.65 | 5.08 | 6.52 | 5.49 | 5.36 |
Recovery
The recovery of Human CDK1 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) | 95-108 | 102 |
| EDTA plasma (n=5) | 85-98 | 90 |
| Cell culture media (n=5) | 93-107 | 100 |
Linearity
Samples were spiked with high concentrations of Human CDK1 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 (%) | 85-95 | 97-108 | 89-105 |
| Average (%) | 90 | 102 | 96 | |
| 1:4 | Range (%) | 88-98 | 80-94 | 85-98 |
| Average (%) | 93 | 86 | 91 | |
| 1:8 | Range (%) | 94-108 | 83-95 | 90-103 |
| Average (%) | 99 | 90 | 95 | |
| 1:16 | Range (%) | 91-101 | 83-96 | 84-99 |
| Average (%) | 96 | 88 | 91 |
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 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.
