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 Human IL1 beta . 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 Human 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 Human IL1 beta, 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 Human IL1 beta. The concentration of Human IL1 beta 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 | Human |
Detection Method | Colormetric |
Detection Range | 7.81-500 pg/mL |
Sensitivity | 4.69 pg/mL |
Sample Volume Required Per Well | 100uL |
Sample Type | Serum, plasma and other biological fluids |
This kit recognizes Human IL1 beta ELISA Kit in samples. No significant cross-reactivity or interference between Human IL1 beta ELISA Kit and analogues was observed.
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 |
---|---|---|---|
500 | 2.387 2.403 | 2.395 | 2.318 |
250 | 1.684 1.706 | 1.695 | 1.618 |
125 | 0.956 0.932 | 0.944 | 0.867 |
62.5 | 0.532 0.538 | 0.535 | 0.458 |
31.25 | 0.299 0.279 | 0.289 | 0.212 |
15.63 | 0.192 0.168 | 0.18 | 0.103 |
7.81 | 0.123 0.137 | 0.13 | 0.053 |
0 | 0.068 0.086 | 0.077 | -- |
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IL1 beta 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 Human IL1 beta ELISA Kit were tested on 3 different plates, 20 replicates in each plate.
Intra-assay Precision | Inter-assay Precision | |||||
---|---|---|---|---|---|---|
Sample | 1 | 2 | 3 | 1 | 2 | 3 |
n | 20 | 20 | 20 | 20 | 20 | 20 |
Mean (pg/mL) | 27.70 | 46.20 | 197.60 | 29.10 | 47.80 | 192.60 |
Standard deviation | 1.40 | 2.60 | 9.50 | 1.70 | 2.80 | 6.00 |
C V (%) | 5.05 | 5.63 | 4.81 | 5.84 | 5.86 | 3.12 |
The recovery of Human IL1 beta 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) | 94-110 | 100 |
EDTA plasma (n=5) | 89-102 | 95 |
Cell culture media (n=5) | 84-98 | 91 |
Samples were spiked with high concentrations of Human IL1 beta 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 (%) | 95-109 | 88-100 | 94-108 |
Average (%) | 102 | 94 | 102 | |
1:4 | Range (%) | 95-109 | 82-94 | 93-105 |
Average (%) | 100 | 88 | 99 | |
1:8 | Range (%) | 98-110 | 82-95 | 97-109 |
Average (%) | 105 | 88 | 103 | |
1:16 | Range (%) | 96-108 | 80-91 | 97-113 |
Average (%) | 102 | 86 | 103 |
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 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 |
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 Chromosomal Location of Human Ortholog: 2q14 Cellular Component: extracellular space; extracellular region; cytosol; 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 nitric oxide biosynthetic process; activation of MAPK activity; positive regulation of transcription, DNA-dependent; positive regulation of interleukin-2 biosynthetic process; germ cell programmed cell death; negative regulation of insulin receptor signaling pathway; positive regulation of lipid catabolic process; positive regulation of NF-kappaB import into nucleus; fever; positive regulation of membrane protein ectodomain proteolysis; response to carbohydrate stimulus; activation of NF-kappaB transcription factor; cell-cell signaling; positive regulation of phagocytosis; positive regulation of T cell mediated immunity; positive regulation of T cell proliferation; neutrophil chemotaxis; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of heterotypic cell-cell adhesion; positive regulation of mitosis; smooth muscle adaptation; interleukin-1 beta production; positive regulation of interleukin-6 production; positive regulation of angiogenesis; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription factor activity; negative regulation of lipid metabolic process; sequestering of triacylglycerol; positive regulation of histone phosphorylation; apoptosis; positive regulation of interleukin-6 biosynthetic process; positive regulation of JNK cascade; signal transduction; positive regulation of vascular endothelial growth factor receptor signaling pathway; positive regulation of protein export from nucleus; positive regulation of interleukin-8 production; negative regulation of cell proliferation; negative regulation of lipid catabolic process; hyaluronan biosynthetic process; protein kinase B signaling cascade; lipopolysaccharide-mediated signaling pathway; inflammatory response; regulation of I-kappaB kinase/NF-kappaB cascade; cytokine and chemokine mediated signaling pathway; MAPKKK cascade; response to ATP; positive regulation of interferon-gamma production; positive regulation of chemokine biosynthetic process; positive regulation of prostaglandin secretion; positive regulation of fever; positive regulation of histone acetylation; immune response; positive regulation of protein amino acid phosphorylation; regulation of insulin secretion; embryo implantation Disease: Gastric Cancer, Hereditary Diffuse |
NCBI Summary: | The protein encoded by this gene is a member of the interleukin 1 cytokine family. This cytokine is produced by activated macrophages as a proprotein, which is proteolytically processed to its active form by caspase 1 (CASP1/ICE). This cytokine is an important mediator of the inflammatory response, and is involved in a variety of cellular activities, including cell proliferation, differentiation, and apoptosis. The induction of cyclooxygenase-2 (PTGS2/COX2) by this cytokine in the central nervous system (CNS) is found to contribute to inflammatory pain hypersensitivity. This gene and eight other interleukin 1 family genes form a cytokine gene cluster on chromosome 2. [provided by RefSeq, Jul 2008] |
UniProt Code: | P01584 |
NCBI GenInfo Identifier: | 62906858 |
NCBI Gene ID: | 3553 |
NCBI Accession: | P01584.2 |
UniProt Secondary Accession: | P01584,Q53X59, Q53XX2, Q7M4S7, Q7RU01, Q96HE5, Q9UCT6 |
UniProt Related Accession: | P01584 |
Molecular Weight: | 30,748 Da |
NCBI Full Name: | Interleukin-1 beta |
NCBI Synonym Full Names: | interleukin 1, beta |
NCBI Official Symbol: | IL1B |
NCBI Official Synonym Symbols: | IL-1; IL1F2; IL1-BETA |
NCBI Protein Information: | interleukin-1 beta; IL-1 beta; catabolin; preinterleukin 1 beta; pro-interleukin-1-beta |
UniProt Protein Name: | Interleukin-1 beta |
UniProt Synonym Protein Names: | Catabolin |
Protein Family: | Interleukin |
UniProt Gene Name: | IL1B |
UniProt Entry Name: | IL1B_HUMAN |