Mouse IL2 ELISA Kit (MOES00661)

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MOES00661
€549

Description

ELISA Kit Technical ManualMSDS

Mouse IL2 (Interleukin 2) ELISA Kit

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 Mouse IL2 . 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 IL2 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 IL2, 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 IL2. The concentration of Mouse IL2 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection MethodColormetric
Detection Range15.63-1000 pg/mL
Sensitivity9.38 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Mouse IL2 ELISA Kit in samples. No significant cross-reactivity or interference between Mouse IL2 ELISA Kit 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
10002.453
2.465
2.4592.381
5001.596
1.622
1.6091.531
2500.964
0.964
0.9640.886
1250.475
0.511
0.4930.415
62.50.272
0.26
0.2660.188
31.250.18
0.18
0.180.102
15.630.126
0.134
0.130.052
00.077
0.079
0.078--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse IL2 ELISA Kit were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse IL2 ELISA Kit were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)49.0087.40385.6052.4082.20365.00
Standard deviation3.303.8020.403.104.8015.70
C V (%)6.734.355.295.925.844.30

Recovery

The recovery of Mouse IL2 ELISA 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)87-10193
EDTA plasma (n=5)88-10394
Cell culture media (n=5)89-10296

Linearity

Samples were spiked with high concentrations of Mouse IL2 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)
1:2Range (%)88-10193-10793-107
Average (%)9510099
1:4Range (%)95-11281-9492-106
Average (%)1038698
1:8Range (%)97-11184-9795-110
Average (%)10589101
1:16Range (%)100-11782-9291-107
Average (%)1088798

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 ELISA 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 Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Mouse IL2 ELISA Kit (MOES00661) 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 IL2 ELISA Kit (MOES00661) Protein Information

UniProt Protein Function:IL2: Produced by T-cells in response to antigenic or mitogenic stimulation, this protein is required for T-cell proliferation and other activities crucial to regulation of the immune response. Can stimulate B-cells, monocytes, lymphokine- activated killer cells, natural killer cells, and glioma cells. A chromosomal aberration involving IL2 is found in a form of T-cell acute lymphoblastic leukemia (T-ALL). Translocation t(4;16)(q26;p13) with involves TNFRSF17. Belongs to the IL-2 family.
UniProt Protein Details:

Protein type:Secreted, signal peptide; Secreted; Oncoprotein; Cytokine

Cellular Component: cytosol; extracellular region; extracellular space

Molecular Function:carbohydrate binding; cytokine activity; glycosphingolipid binding; growth factor activity; interleukin-2 receptor binding; kappa-type opioid receptor binding

Biological Process: adaptive immune response; elevation of cytosolic calcium ion concentration; G-protein coupled receptor protein signaling pathway; immune response; immune system process; negative regulation of B cell apoptosis; negative regulation of heart contraction; negative regulation of inflammatory response; negative regulation of lymphocyte proliferation; negative regulation of protein amino acid phosphorylation; positive regulation of activated T cell proliferation; positive regulation of B cell proliferation; positive regulation of immunoglobulin secretion; positive regulation of interferon-gamma production; positive regulation of interleukin-17 production; positive regulation of isotype switching to IgG isotypes; positive regulation of protein amino acid phosphorylation; positive regulation of regulatory T cell differentiation; positive regulation of T cell differentiation; positive regulation of T cell proliferation; positive regulation of transcription from RNA polymerase II promoter; positive regulation of tyrosine phosphorylation of Stat5 protein; protein kinase C activation; regulation of T cell homeostatic proliferation; response to ethanol

UniProt Code:P04351
NCBI GenInfo Identifier:124326
NCBI Gene ID:16183
NCBI Accession:P04351.1
UniProt Secondary Accession:P04351,P97945, Q791T3,
UniProt Related Accession:P04351
Molecular Weight:19,400 Da
NCBI Full Name:Interleukin-2
NCBI Synonym Full Names:interleukin 2
NCBI Official Symbol:Il2  
NCBI Official Synonym Symbols:Il-2  
NCBI Protein Information:interleukin-2
UniProt Protein Name:Interleukin-2
UniProt Synonym Protein Names:T-cell growth factor; TCGF
Protein Family:Interleukin
UniProt Gene Name:Il2  
UniProt Entry Name:IL2_MOUSE
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Additional Information

Reactivity:
Mouse
ELISA Type:
Sandwich
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