Mouse IL-1 beta/IL-1F2 DuoSet ELISA Best Seller
R&D Systems, part of Bio-Techne | Catalog # DY401
Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
15.6-1000 pg/mL
Sample Type
Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Reactivity
Mouse
Mouse IL-1 beta/IL-1F2 DuoSet ELISA Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Product Summary for Mouse IL-1 beta/IL-1F2 DuoSet ELISA
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse IL-1 beta/IL-1F2. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
100 µL
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Mouse IL-1 beta/IL-1F2 DuoSet ELISA
Mouse IL-1 beta / IL-1F2 ELISA Standard Curve
Detection of Mouse IL-1 beta/IL-1F2 by ELISA
Exosomal release of IL-1 alpha by neutrophils(A) Representative confocal images of peritoneal neutrophils stimulated with LPS or curdlan for 6 h.(B) Quantification of IL-1 alpha and CD63 co-localization using ImageJ (each data point represents a single cell).(C) NTA of EV size distribution and concentration.(D–G) Neutrophils were stimulated in the presence of exosome inhibitor GW4869, and IL-1 alpha and IL-1 beta were quantified by ELISA in isolated EVs following lysis (D and F) and in total cell-free supernatants (E and G).(H) Inhibition of EV secretion shown by NTA.(I) Bioactive IL-1 signaling through IL-1R1 reporter cells was measured in isolated exosomes in the absence of detergent lysis (n = 4). Neutralizing antibodies (Abs) to IL-1 alpha, IL-1 beta, or both cytokines were included in the reporter assay, and bioactive cytokine concentration was calculated based on a standard curve using recombinant IL-1 alpha and IL-1 beta.Two-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Experiments in (A) and (B) were repeated three times; (C)–(G) are biological replicates from repeat experiments. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/34010648), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse IL-1 beta/IL-1F2 by ELISA
Exosomal release of IL-1 alpha by neutrophils(A) Representative confocal images of peritoneal neutrophils stimulated with LPS or curdlan for 6 h.(B) Quantification of IL-1 alpha and CD63 co-localization using ImageJ (each data point represents a single cell).(C) NTA of EV size distribution and concentration.(D–G) Neutrophils were stimulated in the presence of exosome inhibitor GW4869, and IL-1 alpha and IL-1 beta were quantified by ELISA in isolated EVs following lysis (D and F) and in total cell-free supernatants (E and G).(H) Inhibition of EV secretion shown by NTA.(I) Bioactive IL-1 signaling through IL-1R1 reporter cells was measured in isolated exosomes in the absence of detergent lysis (n = 4). Neutralizing antibodies (Abs) to IL-1 alpha, IL-1 beta, or both cytokines were included in the reporter assay, and bioactive cytokine concentration was calculated based on a standard curve using recombinant IL-1 alpha and IL-1 beta.Two-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Experiments in (A) and (B) were repeated three times; (C)–(G) are biological replicates from repeat experiments. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/34010648), licensed under a CC-BY license. Not internally tested by R&D Systems.Kit Contents for Mouse IL-1 beta/IL-1F2 DuoSet ELISA
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-1 beta/IL-1F2
IL-1 alpha and IL-1 beta are structurally related polypeptides that
show approximately 25% homology at the amino acid (aa) level. Both are
synthesized as 31 kDa precursors that are subsequently cleaved into mature
proteins of approximately 17.5 kDa (6, 7). Cleavage of the IL-1 beta precursor by
Caspase-1/ICE is a key step in the inflammatory response (2, 8). Neither IL-1 alpha
nor IL-1 beta contains a typical hydrophobic signal peptide (9-11), but evidence
suggests that these factors can be secreted by non-classical pathways (12, 13).
A portion of unprocessed IL-1 alpha can be presented on the cell membrane and may
retain biological activity (14). The precursor form of IL-1 beta, unlike the IL-1 alpha
precursor, shows little or no biological activity in comparison to the
processed form (13, 15). Both unprocessed and mature forms of IL-1 beta are
exported from the cell.
IL-1 alpha and IL-1 beta exert their effects through immunoglobulin
superfamily receptors that additionally bind IL-1ra. The 80 kDa transmembrane
type I receptor (IL-1 RI) is expressed on T cells, fibroblasts, keratinocytes,
endothelial cells, synovial lining cells, chondrocytes, and hepatocytes (16,
17). The 68 kDa transmembrane type II receptor (IL-1 RII) is expressed on B cells,
neutrophils, and bone marrow cells (18). The two IL-1 receptor types show
approximately 28% homology in their extracellular domains but differ
significantly in that the type II receptor has a cytoplasmic domain of only 29
aa, whereas the type I receptor has a 213 aa cytoplasmic domain. IL-1 RII does
not appear to signal in response to IL-1 and may function as a decoy receptor
that attenuates IL-1 function (19). The IL-1 receptor accessory protein (IL-1
RAcP) associates with IL-1 RI and is required for IL-1 RI signal transduction
(20). IL-1ra is a secreted molecule that functions as a competitive inhibitor
of IL-1 (21, 22). Soluble forms of both IL-1 RI and IL-1 RII have been detected
in human plasma, synovial fluids, and the conditioned media of several human
cell lines (23, 24). In addition, IL-1 binding proteins that resemble soluble
IL-1 RII are encoded by vaccinia and cowpox viruses (25).
Long Name
Interleukin 1 beta
Alternate Names
IL-1b, IL-1F2, IL1 beta, IL1B
Entrez Gene IDs
Gene Symbol
IL1B
Additional IL-1 beta/IL-1F2 Products
Product Documents for Mouse IL-1 beta/IL-1F2 DuoSet ELISA
Product Specific Notices for Mouse IL-1 beta/IL-1F2 DuoSet ELISA
For research use only
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