Rat IL-1 beta CLIA Kit (RTES00012)

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RTES00012
€649

Description

ELISA Kit Technical ManualMSDS

Rat IL1 beta (Interleukin 1 Beta) 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 Rat IL1 beta . 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 Rat IL1 beta 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 Rat IL1 beta, 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 Rat IL1 beta. The concentration of Rat IL1 beta in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityRat
Detection methodChemiluminescence
Detection range12.50-800 pg/mL
Sensitivity7.50 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Rat IL1 beta CLIA Kit in samples. No significant cross-reactivity or interference between Rat IL1 beta 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
80052069
53215
5264252616
40020854
22614
2173421708
20010384
9032
97089682
1004428
4676
45524526
502323
2053
21882162
251110
1010
10601034
12.50497
521
509483
025
27
26--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat IL1 beta 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 Rat IL1 beta CLIA Kit were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)36.4983.36301.9038.3077.17276.82
Standard deviation3.898.9930.104.298.2122.84
C V (%)10.6610.789.9711.2010.648.25

Recovery

The recovery of Rat IL1 beta 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)90-10496
EDTA plasma (n=5)100-115107
Cell culture media (n=5)94-109102

Linearity

Samples were spiked with high concentrations of Rat IL1 beta 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 (%)100-11597-11496-109
Average (%)109104101
1:4Range (%)97-11387-10097-113
Average (%)10393104
1:8Range (%)95-11094-10587-101
Average (%)10110092
1:16Range (%)101-11298-11397-112
Average (%)107106103

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

Rat IL1 beta CLIA Kit (RTES00012) 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.

Rat IL1 beta CLIA Kit (RTES00012) Protein Information

UniProt Protein Function:IL1B: Produced by activated macrophages, IL-1 stimulates thymocyte proliferation by inducing IL-2 release, B-cell maturation and proliferation, and fibroblast growth factor activity. IL-1 proteins are involved in the inflammatory response, being identified as endogenous pyrogens, and are reported to stimulate the release of prostaglandin and collagenase from synovial cells. Monomer. Belongs to the IL-1 family.
UniProt Protein Details:

Protein type:Cytokine

Cellular Component: extracellular space; extracellular region; vesicle; secretory granule

Molecular Function:protein domain specific binding; interleukin-1 receptor binding; cytokine activity

Biological Process: positive regulation of granulocyte macrophage colony-stimulating factor production; negative regulation of MAP kinase activity; positive regulation of JNK activity; positive regulation of nitric oxide biosynthetic process; negative regulation of glutamate secretion; glycoprotein metabolic process; positive regulation of apoptosis; activation of MAPK activity; positive regulation of transcription, DNA-dependent; positive regulation of interleukin-2 biosynthetic process; response to glucocorticoid stimulus; germ cell programmed cell death; negative regulation of insulin receptor signaling pathway; positive regulation of glial cell differentiation; positive regulation of NF-kappaB import into nucleus; response to lipopolysaccharide; positive regulation of lipid catabolic process; fever; positive regulation of membrane protein ectodomain proteolysis; response to organic cyclic substance; response to carbohydrate stimulus; activation of NF-kappaB transcription factor; elevation of cytosolic calcium ion concentration; response to vitamin D; pentacyclic triterpenoid metabolic process; positive regulation of phagocytosis; positive regulation of T cell proliferation; positive regulation of astrocyte differentiation; response to drug; neutrophil chemotaxis; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of heterotypic cell-cell adhesion; positive regulation of mitosis; positive regulation of interleukin-6 production; interleukin-1 beta production; social behavior; response to organic nitrogen; purine base metabolic process; positive regulation of angiogenesis; negative regulation of neuron differentiation; response to ethanol; response to heat; positive regulation of cell division; positive regulation of transcription factor activity; positive regulation of transcription from RNA polymerase II promoter; negative regulation of lipid metabolic process; leukocyte migration; response to peptide hormone stimulus; estrogen metabolic process; sequestering of triacylglycerol; wound healing; positive regulation of interleukin-6 biosynthetic process; response to morphine; positive regulation of JNK cascade; negative regulation of transcription from RNA polymerase II promoter; response to L-ascorbic acid; chronic inflammatory response to antigenic stimulus; positive regulation of stress-activated MAPK cascade; response to estradiol stimulus; negative regulation of neurogenesis; positive regulation of interleukin-8 production; negative regulation of cell proliferation; learning and/or memory; negative regulation of lipid catabolic process; hyaluronan biosynthetic process; protein kinase B signaling cascade; lipopolysaccharide-mediated signaling pathway; response to gamma radiation; regulation of I-kappaB kinase/NF-kappaB cascade; inflammatory response; response to nutrient; aging; cytokine and chemokine mediated signaling pathway; MAPKKK cascade; positive regulation of immature T cell proliferation in the thymus; response to ATP; memory; ovulation; polyketide metabolic process; positive regulation of interferon-gamma production; positive regulation of chemokine biosynthetic process; response to ozone; positive regulation of prostaglandin secretion; response to hypoxia; positive regulation of fever; immune response; positive regulation of protein amino acid phosphorylation; regulation of insulin secretion

NCBI Summary:an inflammatory cytokine [RGD, Feb 2006]
UniProt Code:Q63264
NCBI GenInfo Identifier:2497335
NCBI Gene ID:24494
NCBI Accession:Q63264.1
UniProt Related Accession:Q63264
Molecular Weight:30,644 Da
NCBI Full Name:Interleukin-1 beta
NCBI Synonym Full Names:interleukin 1 beta
NCBI Official Symbol:Il1b  
NCBI Protein Information:interleukin-1 beta; IL-1 beta
UniProt Protein Name:Interleukin-1 beta
Protein Family:Interleukin
UniProt Gene Name:Il1b  
UniProt Entry Name:IL1B_RAT
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Additional Information

Product type:
ELISA
Reactivity:
Rat
ELISA Type:
CLIA
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