Human IL1 beta ELISA Kit (HUES01423)

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HUES01423
€499

Description

ELISA Kit Technical ManualMSDS

Human IL1 beta (Interleukin 1 Beta) 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 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 typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range7.81-500 pg/mL
Sensitivity4.69 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

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.

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
5002.387
2.403
2.3952.318
2501.684
1.706
1.6951.618
1250.956
0.932
0.9440.867
62.50.532
0.538
0.5350.458
31.250.299
0.279
0.2890.212
15.630.192
0.168
0.180.103
7.810.123
0.137
0.130.053
00.068
0.086
0.077--

Precision

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
Sample123123
n202020202020
Mean (pg/mL)27.7046.20197.6029.1047.80192.60
Standard deviation1.402.609.501.702.806.00
C V (%)5.055.634.815.845.863.12

Recovery

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)94-110100
EDTA plasma (n=5)89-10295
Cell culture media (n=5)84-9891

Linearity

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)
1:2Range (%)95-10988-10094-108
Average (%)10294102
1:4Range (%)95-10982-9493-105
Average (%)1008899
1:8Range (%)98-11082-9597-109
Average (%)10588103
1:16Range (%)96-10880-9197-113
Average (%)10286103

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

Human IL1 beta ELISA Kit (HUES01423) 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.

Human IL1 beta ELISA Kit (HUES01423) 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

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
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Additional Information

Reactivity:
Human
ELISA Type:
Sandwich
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