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  1. Link

    human-il-1-beta-elispot-set-ab46578.pdf

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Immunology Innate Immunity Macrophage / Inflamm.
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Human IL-1 beta ELISPOT Set (ab46578)

  • Datasheet
  • SDS
  • Protocol Booklet
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Key features and details

  • Assay type: Sandwich (qualitative)
  • Sample type: Suspension cells
  • Reacts with: Human

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Overview

  • Product name

    Human IL-1 beta ELISPOT Set
    See all IL-1 beta kits
  • Sample type

    Suspension cells
  • Assay type

    Sandwich (qualitative)
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    The Human IL-1 beta ELISPOT Set assay is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing cytokine production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. Elispot assay constitutes an ideal tool in the TH1 / TH2 response, vaccine development, viral infection monitoring and treatment, cancerology, infectious diseases, autoimmune diseases and transplantation.


    This ELISPOT assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.


    Principal:


    After cell stimulation, locally produced cytokines are captured by a specific monoclonal antibody. After cell lysis, trapped cytokine molecules are revealed by a secondary biotinylated detection antibody, which is in turn recognised by streptavidin conjugated to alkaline phosphatase. PVDF-bottomed-well plates are then incubated with BCIP/NBT substrate. Colored "purple" spots indicate cytokine production by individual cells.


    Recognizes natural (pro and mature) human IL-1 beta.

  • Tested applications

    Suitable for: ELISpotmore details
  • Platform

    Reagents

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 5 x 96 tests 10 x 96 tests 15 x 96 tests 20 x 96 tests
    Bovine Serum Albumin 1 x 1g 2 x 1g 3 x 1g 4 x 1g
    Human IL-1β Capture antibody 1 x 500µl 2 x 500µl 3 x 500µl 4 x 500µl
    IL-1ß Biotinylated Detection antibody 1 vial 2 vials 3 vials 4 vials
    Ready-to-use BCIP/NBT substrate buffer 1 x 50ml 2 x 50ml 3 x 50ml 4 x 50ml
    Streptavidin - Alkaline Phosphatase conjugated 1 x 50µl 2 x 50µl 3 x 50µl 4 x 50µl
  • Research areas

    • Immunology
    • Innate Immunity
    • Macrophage / Inflamm.
    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Neuroscience
    • Sensory System
    • Somatosensory system
    • Nociception
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Kits/ Lysates/ Other
    • Kits
    • ELISPOT and FLUOROSPOT Kits
    • ELISPOT Kits
    • Interleukin ELISPOT kits
    • Metabolism
    • Types of disease
    • Obesity
    • Neuroscience
    • Processes
  • 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.
  • Target information above from: UniProt accession P01584 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Catabolin
    • H1
    • IFN beta inducing factor
    • IL 1
    • IL 1 beta
    • IL-1 beta
    • IL1
    • IL1 BETA
    • IL1B
    • IL1B_HUMAN
    • IL1F2
    • Interleukin 1 beta
    • Interleukin 1 beta precursor
    • interleukin 1, beta
    • Interleukin-1 beta
    • OAF
    • Osteoclast activating factor
    • OTTHUMP00000162031
    • Preinterleukin 1 beta
    • Preinterleukin beta
    • Pro interleukin 1 beta
    see all
  • Database links

    • Entrez Gene: 3553 Human
    • Omim: 147720 Human
    • SwissProt: P01584 Human
    • Unigene: 126256 Human

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab46578 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
    ELISpot
    Use at an assay dependent dilution.
    Notes
    ELISpot
    Use at an assay dependent dilution.

    Protocols

    • Human IL1 beta Elispot Kit
    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (1)

    Publishing research using ab46578? Please let us know so that we can cite the reference in this datasheet.

    ab46578 has been referenced in 1 publication.

    • Dissanayake K  et al. Potential applicability of cytokines as biomarkers of disease activity in rheumatoid arthritis: Enzyme-linked immunosorbent spot assay-based evaluation of TNF-a, IL-1ß, IL-10 and IL-17A. PLoS One 16:e0246111 (2021). PubMed: 33497394

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