Human IL24 CLIA Kit (HUES01266)

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HUES01266
€599

Description

ELISA Kit Technical ManualMSDS

Human IL24 (Interleukin 24) 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 IL24 . 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 IL24 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 IL24, 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 IL24. The concentration of Human IL24 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 range7.81-500 pg/mL
Sensitivity4.69 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Human IL24 CLIA Kit in samples. No significant cross-reactivity or interference between Human IL24 CLIA Kit 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
50050607
52467
5153751508
25018974
23022
2099820969
1259805
8987
93969367
62.54442
4582
45124483
31.252455
2143
22992270
15.631286
1214
12501221
7.81715
765
740711
028
30
29--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IL24 CLIA Kit were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IL24 CLIA Kit were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)26.2658.64204.1028.7254.72221.22
Standard deviation2.246.2414.252.314.0616.41
C V (%)8.5310.646.988.047.427.42

Recovery

The recovery of Human IL24 CLIA Kit 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)87-10092
EDTA plasma (n=5)94-109100
Cell culture media (n=5)89-9994

Linearity

Samples were spiked with high concentrations of Human IL24 CLIA Kit 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 (%)91-10490-10298-114
Average (%)9895105
1:4Range (%)94-10586-9884-96
Average (%)999191
1:8Range (%)86-9788-10399-114
Average (%)9294104
1:16Range (%)98-112103-11685-97
Average (%)10610892

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 IL24 CLIA Kit (HUES01266) 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 IL24 CLIA Kit (HUES01266) Protein Information

UniProt Protein Function:IL24: Has antiproliferative properties on melanoma cells and may contribute to terminal cell differentiation. Belongs to the IL-10 family. 4 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Motility/polarity/chemotaxis; Secreted; Secreted, signal peptide

Chromosomal Location of Human Ortholog: 1q32

Cellular Component: extracellular space; endoplasmic reticulum

Molecular Function:cytokine activity

Biological Process: negative regulation of cell proliferation; wound healing; apoptosis; positive regulation of cell proliferation; positive regulation of JAK-STAT cascade; positive regulation of caspase activity; immune response; negative regulation of caspase activity; inflammatory response; negative regulation of cell migration; positive regulation of tyrosine phosphorylation of Stat3 protein; serine phosphorylation of STAT3 protein

NCBI Summary:This gene encodes a member of the IL10 family of cytokines. It was identified as a gene induced during terminal differentiation in melanoma cells. The protein encoded by this gene can induce apoptosis selectively in various cancer cells. Overexpression of this gene leads to elevated expression of several GADD family genes, which correlates with the induction of apoptosis. The phosphorylation of mitogen-activated protein kinase 14 (MAPK7/P38), and heat shock 27kDa protein 1 (HSPB2/HSP27) are found to be induced by this gene in melanoma cells, but not in normal immortal melanocytes. Alternatively spliced transcript variants encoding distinct isoforms have been reported. [provided by RefSeq, Jul 2008]
UniProt Code:Q13007
NCBI GenInfo Identifier:2497340
NCBI Gene ID:11009
NCBI Accession:Q13007.1
UniProt Related Accession:Q13007
Molecular Weight:
NCBI Full Name:Interleukin-24
NCBI Synonym Full Names:interleukin 24
NCBI Official Symbol:IL24  
NCBI Official Synonym Symbols:C49A; FISP; MDA7; MOB5; ST16; IL10B  
NCBI Protein Information:interleukin-24
UniProt Protein Name:Interleukin-24
UniProt Synonym Protein Names:Melanoma differentiation-associated gene 7 protein; MDA-7; Suppression of tumorigenicity 16 protein
Protein Family:Interleukin
UniProt Gene Name:IL24  
UniProt Entry Name:IL24_HUMAN
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Additional Information

Reactivity:
Human
ELISA Type:
CLIA
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