Mouse IL-6 ELISA Kit (MOES00663)

SKU:
MOES00663
€299

Description

ELISA Kit Technical ManualMSDS

Mouse IL6 (Interleukin 6) 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 IL6 . 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 IL6 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 IL6, 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 IL6. The concentration of Mouse IL6 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 Range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Mouse IL6 ELISA Kit in samples. No significant cross-reactivity or interference between Mouse IL6 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
20002.471
2.503
2.4872.427
10001.682
1.72
1.7011.641
5000.945
0.915
0.930.87
2500.421
0.441
0.4310.371
1250.267
0.243
0.2550.195
62.50.17
0.154
0.1620.102
31.250.109
0.115
0.1120.052
00.05
0.07
0.06--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse IL6 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 IL6 ELISA Kit were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)99.40174.50952.80105.30162.701040.70
Standard deviation6.407.7050.506.508.1038.50
C V (%)6.444.415.306.174.983.70

Recovery

The recovery of Mouse IL6 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)86-10093
EDTA plasma (n=5)88-10194
Cell culture media (n=5)89-10495

Linearity

Samples were spiked with high concentrations of Mouse IL6 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 (%)91-10691-10398-111
Average (%)9898104
1:4Range (%)94-10988-10398-110
Average (%)10094104
1:8Range (%)94-10887-9895-106
Average (%)10193100
1:16Range (%)97-11384-9797-109
Average (%)10591103

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 IL6 ELISA Kit (MOES00663) 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 IL6 ELISA Kit (MOES00663) Protein Information

UniProt Protein Function:IL6: Cytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Plays an essential role in the final differentiation of B-cells into Ig- secreting cells Involved in lymphocyte and monocyte differentiation. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation Acts on B-cells, T-cells, hepatocytes, hematopoietic progenitor cells and cells of the CNS. Also acts as a myokine. It is discharged into the bloodstream after muscle contraction and acts to increase the breakdown of fats and to improve insulin resistance. Genetic variations in IL6 are associated with susceptibility to rheumatoid arthritis systemic juvenile (RASJ). An inflammatory articular disorder with systemic- onset beginning before the age of 16. It represents a subgroup of juvenile arthritis associated with severe extraarticular features and occasionally fatal complications. During active phases of the disorder, patients display a typical daily spiking fever, an evanescent macular rash, lymphadenopathy, hepatosplenomegaly, serositis, myalgia and arthritis. A IL6 promoter polymorphism is associated with a lifetime risk of development of Kaposi sarcoma in HIV-infected men. Belongs to the IL-6 superfamily.
UniProt Protein Details:

Protein type:Secreted, signal peptide; Secreted

Cellular Component: extracellular space; cell; cytoplasm; extracellular region; interleukin-6 receptor complex; external side of plasma membrane

Molecular Function:protein binding; interleukin-6 receptor binding; growth factor activity; cytokine activity; receptor binding

Biological Process: positive regulation of nitric oxide biosynthetic process; positive regulation of apoptosis; negative regulation of collagen biosynthetic process; positive regulation of transcription, DNA-dependent; response to glucocorticoid stimulus; negative regulation of cytokine secretion; myeloid cell homeostasis; positive regulation of JAK-STAT cascade; glucose homeostasis; positive regulation of tyrosine phosphorylation of Stat3 protein; muscle maintenance; regulation of apoptosis; regulation of cell shape; positive regulation of T-helper 2 cell differentiation; positive regulation of acute inflammatory response; negative regulation of gluconeogenesis; acute-phase response; positive regulation of T cell proliferation; cell growth; defense response to virus; neurite development; positive regulation of protein import into nucleus, translocation; positive regulation of interleukin-6 production; defense response to protozoan; positive regulation of chemokine production; positive regulation of peptidyl-tyrosine phosphorylation; cell redox homeostasis; positive regulation of B cell activation; negative regulation of protein kinase activity; neutrophil apoptosis; positive regulation of transcription factor activity; positive regulation of transcription from RNA polymerase II promoter; positive regulation of epithelial cell proliferation; negative regulation of apoptosis; negative regulation of muscle development; T cell activation; positive regulation of translation; negative regulation of hormone secretion; positive regulation of smooth muscle cell proliferation; negative regulation of caspase activity; regulation of circadian sleep/wake cycle, non-REM sleep; negative regulation of cell proliferation; positive regulation of MAPKKK cascade; positive regulation of cell proliferation; response to wounding; hepatic immune response; inflammatory response; negative regulation of chemokine biosynthetic process; cytokine and chemokine mediated signaling pathway; positive regulation of immunoglobulin secretion; positive regulation of peptidyl-serine phosphorylation; endocrine pancreas development; regulation of cell proliferation; positive regulation of protein kinase B signaling cascade; immune response; positive regulation of neuron differentiation; positive regulation of DNA replication; positive regulation of transmission of nerve impulse

UniProt Code:P08505
NCBI GenInfo Identifier:124348
NCBI Gene ID:16193
NCBI Accession:P08505.1
UniProt Secondary Accession:P08505,Q3UCQ0, Q8BN26,
UniProt Related Accession:P08505
Molecular Weight:24,384 Da
NCBI Full Name:Interleukin-6
NCBI Synonym Full Names:interleukin 6
NCBI Official Symbol:Il6
NCBI Official Synonym Symbols:Il-6
NCBI Protein Information:interleukin-6; interleukin HP-1; B-cell hybridoma growth factor
UniProt Protein Name:Interleukin-6
UniProt Synonym Protein Names:B-cell hybridoma growth factor; Interleukin HP-1
Protein Family:Interleukin
UniProt Gene Name:Il6
UniProt Entry Name:IL6_MOUSE
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Additional Information

Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
Sandwich
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