Human DCLK1 (Doublecortin Like Kinase 1) CLIA Kit (HUES00262)

SKU:
HUES00262
€599

Description

ELISA Kit Technical ManualMSDS

Human DCLK1 (Doublecortin Like Kinase 1) CLIA Kit

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 DCLK1 . 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 DCLK1 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 DCLK1, 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 DCLK1. The concentration of Human DCLK1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection methodChemiluminescence
Detection range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Human DCLK1 in samples. No significant cross-reactivity or interference between Human DCLK1 and analogues was observed.

Typical Data

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.

Concentration
(pg/mL)
RLU Average Corrected
200052838
58604
5572155693
100022127
25821
2397423946
50011631
10433
1103211004
2505040
5548
52945266
1252701
2515
26082580
62.51337
1285
13111283
31.25669
679
674646
028
28
28--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human DCLK1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human DCLK1 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)109.40211.13695.61119.91221.25630.25
Standard deviation11.8618.9053.2110.0523.7238.57
C V (%)10.848.957.658.3810.726.12

Recovery

The recovery of Human DCLK1 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)91-10798
EDTA plasma (n=5)87-9892
Cell culture media (n=5)91-10497

Linearity

Samples were spiked with high concentrations of Human DCLK1 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)
1:2Range (%)92-10798-11294-108
Average (%)9810699
1:4Range (%)85-9792-10788-100
Average (%)929893
1:8Range (%)100-11898-10989-102
Average (%)10910495
1:16Range (%)99-11397-109100-114
Average (%)104103108

Kit Components & Storage

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 CLIA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human DCLK1 (Doublecortin Like Kinase 1) CLIA Kit (HUES00262) Assay procedure

    1. 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.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. 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.
    7. 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.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. 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.
    11. Determine the RLU value of each well immediately.

Human DCLK1 (Doublecortin Like Kinase 1) CLIA Kit (HUES00262) Protein Information

UniProt Protein Function:Probable kinase that may be involved in a calcium-signaling pathway controlling neuronal migration in the developing brain. May also participate in functions of the mature nervous system.
NCBI Summary:This gene encodes a member of the protein kinase superfamily and the doublecortin family. The protein encoded by this gene contains two N-terminal doublecortin domains, which bind microtubules and regulate microtubule polymerization, a C-terminal serine/threonine protein kinase domain, which shows substantial homology to Ca2+/calmodulin-dependent protein kinase, and a serine/proline-rich domain in between the doublecortin and the protein kinase domains, which mediates multiple protein-protein interactions. The microtubule-polymerizing activity of the encoded protein is independent of its protein kinase activity. The encoded protein is involved in several different cellular processes, including neuronal migration, retrograde transport, neuronal apoptosis and neurogenesis. This gene is up-regulated by brain-derived neurotrophic factor and associated with memory and general cognitive abilities. Multiple transcript variants generated by two alternative promoter usage and alternative splicing have been reported, but the full-length nature and biological validity of some variants have not been defined. These variants encode different isoforms, which are differentially expressed and have different kinase activities.[provided by RefSeq, Sep 2010]
UniProt Code:O15075
NCBI GenInfo Identifier:6225242
NCBI Gene ID:9201
NCBI Accession:O15075.2
UniProt Secondary Accession:O15075,Q5VZY8, Q5VZZ0, Q5VZZ1, B7Z3E9,
UniProt Related Accession:O15075
Molecular Weight:47,681 Da
NCBI Full Name:Serine/threonine-protein kinase DCLK1
NCBI Synonym Full Names:doublecortin like kinase 1
NCBI Official Symbol:DCLK1
NCBI Official Synonym Symbols:CL1; DCLK; CLICK1; DCDC3A; DCAMKL1
NCBI Protein Information:serine/threonine-protein kinase DCLK1
UniProt Protein Name:Serine/threonine-protein kinase DCLK1
UniProt Synonym Protein Names:Doublecortin domain-containing protein 3A; Doublecortin-like and CAM kinase-like 1; Doublecortin-like kinase 1
Protein Family:Serine/threonine-protein kinase
UniProt Gene Name:DCLK1
UniProt Entry Name:DCLK1_HUMAN
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Additional Information

Product type:
ELISA
Reactivity:
Human
ELISA Type:
CLIA
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