Overview

  • Product name
    Mouse IL-1 beta ELISA Kit, Fluorescent
    See all IL-1 beta kits
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Cell medium 5 6.1%
    Inter-assay
    Sample n Mean SD CV%
    Cell medium 3 7.7%
  • Sample type
    Cell culture supernatant, Urine, Serum, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    0.2 pg/ml
  • Range
    0.2 pg/ml - 200 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 105 100% - 108%
    Urine 94 91% - 95%
    Serum 101 98% - 103%
    Plasma 89 78% - 95%
    Tissue Homogenate 97 95% - 100%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Mouse
    Does not react with: Rabbit, Hamster, Dog, Human
  • Product overview

    IL-1 beta (Interleukin-1 beta) in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-1 beta (Interleukin-1 beta) protein in mouse serum, plasma, urine, cell culture supernatant, and cell and tissue extracts.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material.  CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • Notes

    Interleukin-1 beta (IL-1 beta) is a 17.5 kDa cytokine protein of the Interleukin 1 family. IL-1 beta is secreted by macrophages and other cell types in response to inflammatory agents or infection. IL-1 beta plays a role in a number of cellular processes, including immune responses, bone remodeling, apoptosis and inflammatory pain hypersensitivity.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Stoplight Red Substrate 1 x 120µl
    10X Mouse IL-1 beta Capture Antibody 1 x 600µl
    10X Mouse IL-1 beta Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    50X Cell Extraction Enhancer Solution 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 5BI 1 x 6ml
    Mouse IL-1 beta Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • Research areas
  • Function
    Potent proinflammatory cytokine. Initially discovered as the major endogenous pyrogen, induces prostaglandin synthesis, neutrophil influx and activation, T-cell activation and cytokine production, B-cell activation and antibody production, and fibroblast proliferation and collagen production. Promotes Th17 differentiation of T-cells.
  • Tissue specificity
    Expressed in activated monocytes/macrophages (at protein level).
  • Sequence similarities
    Belongs to the IL-1 family.
  • Post-translational
    modifications
    Activation of the IL1B precursor involves a CASP1-catalyzed proteolytic cleavage. Processing and secretion are temporarily associated.
  • Cellular localization
    Cytoplasm, cytosol. Lysosome. Secreted, exosome. Cytoplasmic vesicle, autophagosome. Secreted. The precursor is cytosolic. In response to inflammasome-activating signals, such as ATP for NLRP3 inflammasome or bacterial flagellin for NLRC4 inflammasome, cleaved and secreted. IL1B lacks any known signal sequence and the pathway(s) of its secretion is(are) not yet fully understood (PubMed:24201029). On the basis of experimental results, several unconventional secretion mechanisms have been proposed. 1. Secretion via secretory lysosomes: a fraction of CASP1 and IL1B precursor may be incorporated, by a yet undefined mechanism, into secretory lysosomes that undergo Ca(2+)-dependent exocytosis with release of mature IL1B (PubMed:15192144). 2. Secretory autophagy: IL1B-containing autophagosomes may fuse with endosomes or multivesicular bodies (MVBs) and then merge with the plasma membrane releasing soluble IL1B or IL1B-containing exosomes (PubMed:24201029). However, autophagy impacts IL1B production at several levels and its role in secretion is still controversial. 3. Secretion via exosomes: ATP-activation of P2RX7 leads to the formation of MVBs containing exosomes with entrapped IL1B, CASP1 and other inflammasome components. These MVBs undergo exocytosis with the release of exosomes. The release of soluble IL1B occurs after the lysis of exosome membranes (By similarity). 4. Secretion by microvesicle shedding: activation of the ATP receptor P2RX7 may induce an immediate shedding of membrane-derived microvesicles containing IL1B and possibly inflammasome components. The cytokine is then released in the extracellular compartment after microvesicle lysis (PubMed:11728343). 5. Release by translocation through permeabilized plasma membrane. This may occur in cells undergoing pyroptosis due to sustained activation of the inflammasome (By similarity). These mechanisms may not be not mutually exclusive.
  • Information by UniProt
  • Alternative names
    • Catabolin
    • H1
    • IL 1
    • IL 1 beta
    • IL-1 beta
    • IL1 BETA
    • IL1B
    • IL1B_HUMAN
    • IL1F2
    • Interleukin 1 beta
    • Interleukin-1 beta
    • OAF
    • OTTHUMP00000162031
    • Preinterleukin 1 beta
    • Pro interleukin 1 beta
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab229440 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • RAW264.7 cells were grown in the absence (unstimulated) or presence of 5 µg/mL Lipopolysaccharide (LPS) (stimulated) for 48 hours. IL-1 beta was measured in 2-fold diluted cell culture supernatants of unstimulated and LPS stimulated RAW264.7 and cell culture media. Measured values were interpolated from the IL-1 beta Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed: 1.1 pg/mL unstimulated, 83.5 pg/mL stimulated. There was no detectable signal in media.

  • Demonstration of the linearity of dilution by the titration of RAW264.7 stimulated for 48 hours with LPS undiluted to 32-fold dilution in Sample Diluent NS. Background-subtracted data values (mean +/- SD, n = 2) are graphed.

Protocols

References

ab229440 has not yet been referenced specifically in any publications.

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