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Human CDK5 ELISA Kit

SKU:
HUFI00785
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q00535
Sensitivity:
0.188ng/ml
Range:
0.313-20ng/ml
ELISA Type:
Sandwich
Synonyms:
CDK5, Cyclin-dependent kinase 5, TPKII catalytic subunit, Tau protein kinase II catalytic subunit, Cell division protein kinase 5, Serine, threonine-protein kinase PSSALRE
Reactivity:
Human
$719
Frequently bought together:

Description

Human CDK5 ELISA Kit

The Human CDK5 ELISA Kit is a powerful tool for researchers looking to accurately measure CDK5 levels in human samples such as serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures reliable and reproducible results for a variety of research applications.CDK5, or Cyclin-dependent kinase 5, is a critical protein kinase involved in cell cycle regulation, neuronal development, and neurodegenerative diseases. By quantifying CDK5 levels, researchers can gain valuable insights into the role of this protein in various physiological processes and disease states, including Alzheimer's disease and cancer.

This ELISA kit provides a user-friendly and efficient method for measuring CDK5 levels, making it an indispensable tool for studying the molecular mechanisms underlying disease pathology and identifying potential therapeutic targets. Elevate your research with the Human CDK5 ELISA Kit from Assay Genie.

Product Name:Human CDK5 ELISA Kit
Product Code:HUFI00785
Size:96 Assays
Alias:CDK5, Cyclin-dependent kinase 5, TPKII catalytic subunit, Tau protein kinase II catalytic subunit, Cell division protein kinase 5, Serine, threonine-protein kinase PSSALRE
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human CDK5 concentrations in serum plasma and other biological fluids.
Sensitivity:0.188ng/ml
Range:0.313-20ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human CDK5 and the recovery rates were calculated by comparing the measured value to the expected amount of Human CDK5 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)88-10598
EDTA plasma(n=5)88-10495
UFH plasma(n=5)88-10595
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human CDK5 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)87-105%91-103%87-96%
EDTA plasma(n=5)85-96%85-99%82-101%
UFH plasma(n=5)93-98%80-100%91-100%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotQ00535
UniProt Protein Function:CDK5: a protein kinase of the CDK family. Unlike other members of this family, it is not activated by cyclins but by p35 (CDK5R1) and p39. An important regulator of neuronal positioning during brain development. May also play a role in synaptogenesis and neurotransmission. Substrates include TAU, MAP2, NF-H and -M, Nudel, PDE6, beta-catenin, amphyphysin, dynamin I, synapsin 1, Munc-18, and NMDA receptor 2A. Plays a role in myogenesis, haematopoietic cell differentiation, spermatogenesis, insulin secretion, and lens differentiation. Implicated in the pathology of neurofibrillary tangles and formation of senile plaques, hallmarks of Alzheimer?s disease. Induces tau phosphorylation and aggregation and neurofibrillary tangle deposition and neurodegeneration in in vitro and in vivo animal models. Brain samples from Alzeimer?s pateints show elevated CDK5 activity.
UniProt Protein Details:Protein type:Protein kinase, CMGC; Kinase, protein; Cell cycle regulation; Protein kinase, Ser/Thr (non-receptor); EC 2.7.11.1; CMGC group; CDK family; CDK5 subfamily; CDK/CDK5 subfamily

Chromosomal Location of Human Ortholog: 7q36

Cellular Component: axon; cell junction; cell soma; cytoplasm; cytosol; dendrite; growth cone; membrane; neuromuscular junction; nucleoplasm; nucleus; plasma membrane; postsynaptic density

Molecular Function:acetylcholine receptor activator activity; cyclin-dependent protein kinase activity; ErbB-2 class receptor binding; ErbB-3 class receptor binding; kinase activity; protein binding; protein kinase activity; protein serine/threonine kinase activity; tau-protein kinase activity

Biological Process: axon extension; cell proliferation; negative regulation of proteolysis; negative regulation of transcription, DNA-dependent; neurite development; neuron apoptosis; neuron differentiation; neuron migration; oligodendrocyte differentiation; peptidyl-serine phosphorylation; phosphorylation; positive regulation of neuron apoptosis; regulation of apoptosis; regulation of macroautophagy; regulation of synaptic plasticity; synaptic transmission; synaptic vesicle endocytosis; synaptic vesicle exocytosis; synaptogenesis

Disease: Lissencephaly 7 With Cerebellar Hypoplasia

NCBI Summary:This gene encodes a proline-directed serine/threonine kinase that is a member of the cyclin-dependent kinase family of proteins. Unlike other members of the family, the protein encoded by this gene does not directly control cell cycle regulation. Instead the protein, which is predominantly expressed at high levels in mammalian postmitotic central nervous system neurons, functions in diverse processes such as synaptic plasticity and neuronal migration through phosphorylation of proteins required for cytoskeletal organization, endocytosis and exocytosis, and apoptosis. In humans, an allelic variant of the gene that results in undetectable levels of the protein has been associated with lethal autosomal recessive lissencephaly-7. Alternative splicing results in multiple transcript variants. [provided by RefSeq, May 2015]
UniProt Code:Q00535
NCBI GenInfo Identifier:4033704
NCBI Gene ID:1020
NCBI Accession:Q00535.3
UniProt Secondary Accession:Q00535,A1XKG3,
UniProt Related Accession:Q00535
Molecular Weight:29,544 Da
NCBI Full Name:Cyclin-dependent-like kinase 5
NCBI Synonym Full Names:cyclin dependent kinase 5
NCBI Official Symbol:CDK5  
NCBI Official Synonym Symbols:LIS7; PSSALRE  
NCBI Protein Information:cyclin-dependent-like kinase 5
UniProt Protein Name:Cyclin-dependent-like kinase 5
UniProt Synonym Protein Names:Cell division protein kinase 5; Serine/threonine-protein kinase PSSALRE; Tau protein kinase II catalytic subunit; TPKII catalytic subunit
Protein Family:CDK5RAP1-like protein
UniProt Gene Name:CDK5  
UniProt Entry Name:CDK5_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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