Mouse IL-18 ELISA Kit (MOES01225)

SKU:
MOES01225
€299

Description

ELISA Kit Technical ManualMSDS

Mouse IL-18 ELISA Kit

This Mouse IL-18 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 Mouse IL18 . 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 Mouse IL18 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 Mouse IL18, 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 Mouse IL18. The concentration of Mouse IL18 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay type Sandwich
Format 96T
Assay time 4.5h
Reactivity Mouse
Detection Method Colormetric
Detection Range 15.63-1000 pg/mL
Sensitivity 9.38 pg/mL
Sample Volume Required Per Well 100uL
Sample Type Serum, plasma and other biological fluids

Specificity

This Mouse IL-18 kit recognizes IL-18 in samples. No significant cross-reactivity or interference between Mouse IL18 ELISA and analogues was observed.

Typical Data

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
(pg/mL)

O.D

Average

Corrected
1000 2.423
2.479
2.451 2.39
500 1.697
1.743
1.72 1.659
250 0.945
0.921
0.933 0.872
125 0.444
0.462
0.453 0.392
62.5 0.24
0.234
0.237 0.176
31.25 0.165
0.163
0.164 0.103
15.63 0.105
0.123
0.114 0.053
0 0.056
0.066
0.061 --

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse IL-18 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse IL-18 were tested on 3 different plates, 20 replicates in each plate.

 Intra-assay PrecisionInter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 49.50 145.20 342.80 53.00 144.60 363.40
Standard deviation 2.60 8.10 16.80 3.70 7.40 12.40
C V (%) 5.25 5.58 4.90 6.98 5.12 3.41

Recovery

The recovery of Mouse IL-18 ELISA Kit spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample TypeRange (%)Average Recovery (%)
Serum (n=5) 91-102 97
EDTA plasma (n=5) 86-100 91
Cell culture media (n=5) 92-104 99

Linearity

Samples were spiked with high concentrations of Mouse IL18 ELISA 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)
1:2 Range (%) 91-105 87-101 86-99
Average (%) 97 92 93
1:4 Range (%) 97-109 81-93 94-110
Average (%) 104 87 101
1:8 Range (%) 96-109 80-95 97-110
Average (%) 101 87 103
1:16 Range (%) 96-109 88-99 96-112
Average (%) 101 93 104

Mouse IL-18 ELISA 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.

ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 8 wells X12 strips -20'C, 6 months
Reference Standard 2 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 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 (25X) 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

Mouse IL-18 ELISA Kit (MOES01225) 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 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.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Mouse IL18 ELISA Kit (MOES01225) Protein Information

UniProt Protein Function: IL18: Augments natural killer cell activity in spleen cells and stimulates interferon gamma production in T-helper type I cells. Belongs to the IL-1 family.
UniProt Protein Details:

Protein type:Cytokine

Cellular Component: extracellular space; apical plasma membrane; cytoplasm; extracellular region

Molecular Function:cytokine activity

Biological Process: positive regulation of granulocyte macrophage colony-stimulating factor production; positive regulation of apoptosis; positive regulation of smooth muscle cell proliferation; positive regulation of collagen biosynthetic process; positive regulation of NK T cell proliferation; natural killer cell activation; positive regulation of NF-kappaB import into nucleus; positive regulation of smooth muscle cell migration; positive regulation of activated T cell proliferation; positive regulation of interleukin-8 production; positive regulation of superoxide release; positive regulation of neuron apoptosis; lipopolysaccharide-mediated signaling pathway; angiogenesis; inflammatory response; positive regulation of interleukin-17 production; regulation of cell adhesion; negative regulation of peptidyl-tyrosine phosphorylation; positive regulation of natural killer cell proliferation; T-helper 1 type immune response; MAPKKK cascade; interleukin-13 biosynthetic process; positive regulation of tumor necrosis factor production; sleep; positive regulation of chemokine production; positive regulation of vascular permeability; negative regulation of neutrophil apoptosis; detection of mechanical stimulus involved in sensory perception of pain; positive regulation of interferon-gamma production; induction of apoptosis via death domain receptors; response to hypoxia; interferon-gamma biosynthetic process; immune response; negative regulation of myoblast differentiation

UniProt Code: P70380
NCBI GenInfo Identifier: 3219810
NCBI Gene ID: 16173
NCBI Accession: P70380.2
UniProt Related Accession: P70380
Molecular Weight:  
NCBI Full Name: Interleukin-18
NCBI Synonym Full Names: interleukin 18
NCBI Official Symbol: Il18  
NCBI Official Synonym Symbols: Igif; Il-18  
NCBI Protein Information: interleukin-18; IL-1 gamma; interleukin-1 gamma; IFN-gamma-inducing factor; interferon gamma-inducing factor; interferon-gamma-inducing factor
UniProt Protein Name: Interleukin-18
UniProt Synonym Protein Names: Interferon gamma-inducing factor; IFN-gamma-inducing factor; Interleukin-1 gamma; IL-1 gamma
Protein Family: Interleukin
UniProt Gene Name: Il18  
UniProt Entry Name: IL18_MOUSE
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Additional Information

Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
Sandwich
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