Overview

  • Product name
    Human IL-1 beta ELISA Kit
    See all IL1 beta kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 8 4.8%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 5.6%
  • Sample type
    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    5.64 pg/ml
  • Range
    14.06 pg/ml - 900 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 98 96% - 100%
    Serum 103 101% - 105%
    Heparin Plasma 100 99% - 101%
    EDTA Plasma 93 90% - 96%
    Citrate Plasma 86 84% - 88%

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

    Abcam’s Human IL-1 beta SimpleStep ELISA® kit (ab184861) has been re-developed with new capture and detector antibodies.  This new kit will have the same name but a different product number (ab214025). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA platform that provide a higher sensitivity when quantification of IL-1 beta in human serum, plasma, and cell culture supernatants.


    IL-1beta in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-1beta protein in human serum, plasma and cell culture supernatants.


    The 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. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Interleukin 1 beta (IL-1 beta) is produced by activated macrophages and 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.

  • 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
    10X Human IL-1beta Capture Antibody 1 x 600µl
    10X Human IL-1beta Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BI 1 x 6ml
    Human IL-1beta Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Sample Diluent NS 1 x 50ml
    Stop Solution 1 x 12ml
    TMB Substrate 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

Associated products

Applications

Our Abpromise guarantee covers the use of ab214025 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

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

  • The concentrations of IL-1beta were measured in duplicates, interpolated from the IL-1beta standard curves and corrected for sample dilution. Undiluted samples are as follows: PBMC supernatant 50% and 100% THP-1 supernatant. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-1beta concentration was determined to be 1301 pg/mL in PBMC supernatant and 734 pg/mL in THP-1 supernatant.

  • Interpolated values are plotted (mean +/- SD, n=2). IL-1beta was measured in 2 donor serum samples (30 pg/mL and 140 pg/mL) and the remaining 8 samples measured less than the lowest point of the IL-1beta standard curve.

  • Conditioned media was harvested after 48 hours. IL-1beta was measured in 50% unstimulated and PHA stimulated PBMC supernatant. The concentrations of IL-1beta were measured in duplicate, interpolated from the IL-1beta standard curves and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-1beta concentration was determined to be 1273 pg/mL in PHA stimulated PBMC supernatant. There was no detectable signal in unstimulated supernatant.

  • Conditioned media was harvested after 48 hours. IL-1beta was measured in 100% unstimulated and LPS stimulated THP-1 supernatant. The concentrations of IL-1beta were measured in duplicate and interpolated from the IL-1beta standard curves. The interpolated values are plotted (mean +/- SD, n=2). The mean IL-1beta concentration was determined to be 718 pg/mL in LPS stimulated THP-1 supernatant. There was no detectable signal in unstimulated supernatant.

Protocols

References

ab214025 has not yet been referenced specifically in any publications.

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