Overview

  • Product name

    Mouse IL-1 beta ELISA kit
    See all IL-1 beta kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 4.3%
    Inter-assay
    Sample n Mean SD CV%
    Overall 7.5%
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    = 0.016 ng/ml
  • Range

    0.016 ng/ml - 1 ng/ml
  • Recovery

    98 %

  • Assay time

    5h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Abcam’s IL-1 beta (Interleukin-1 beta) mouse in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse IL-1 beta concentrations in plasma, serum and cell culture supernatants.


    An IL-1 beta specific antibody has been precoated onto 96-well plates and blocked. Standards or test samples are added to the wells and subsequently an IL-1 beta specific biotinylated detection antibody is added and then followed by washing with wash buffer. Streptavidin-Peroxidase Conjugate is added and unbound conjugates are washed away with wash buffer. TMB is then used to visualize Streptavidin-Peroxidase enzymatic reaction. TMB is catalyzed by Streptavidin-Peroxidase to produce a blue color product that changes into yellow after adding acidic stop solution. The density of yellow coloration is directly proportional to the amount of IL-1 beta captured in plate.


    Get better reproducibility in only 90 minutes with Mouse IL-1 beta ELISA Kit (ab197742) from our SimpleStep ELISA® range.


    The entire kit may be stored at -20°C for long term storage before reconstitution - Avoid repeated freeze-thaw cycles.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 x 96 tests
    100X Streptavidin-Peroxidase Conjugate 1 x 80µl
    10X Diluent N Concentrate 1 x 30ml
    20X Wash Buffer Concentrate 2 x 30ml
    50X Biotinylated Mouse IL-1 beta Antibody 1 x 120µl
    Chromogen Substrate 1 x 8ml
    IL-1 beta Microplate (12 x 8 well strips) 1 unit
    IL-1 beta Standard 1 vial
    Sealing Tapes 3 units
    Stop Solution 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
    • 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

Associated products

Applications

Our Abpromise guarantee covers the use of ab108866 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 dilution.

Images

  • Representative Standard Curve using ab108866

Protocols

References

ab108866 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Gracias por contactarnos.


He querido comprobar la posibilidad de presentar reactividad cruzada entre ratón y humano en los kits contra las citoquinas que mencionas.

Tenemos varios kits contra dichas citoquinas específicos para ratón, y específicos para humano. Después de estudiar la homología entre secuencias, me temo que no podemos predecir que exista reactividad cruzada entre especies. Experimentalmente tampoco tenemos datos para confirmarlo.

Siento no poder ser de mas ayuda. No dudes en contactarnos para cualquier otra consulta.

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Answer

Thank you for your reply.


Because we carry over 100,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.


Regrettably, as we haven't tested or even compared the reactivity to the pro- or mature form of the IL1 beta, we cannot guarantee it will work with both forms.

However, we are still waiting for a response from our collaborator of ab108866. So there is still a change that the reactivity has been measured.


I hope this information is still helpful. We will let you know as soon as we got a reply.

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Answer

Thank you for contacting us. Please be advised that we have not done any test with p-NPP substrate so we will not be able to provide any data regarding this. We have only tested this Kit using the reagent mentioned on the KIT datasheet. Regarding the standard curve customers have to prepare standard curve by themselves. We have provided everything in the KIT for the preparation of standard curve. Please check in literature what standard curve really mean and then prepare it following the protocol attached to the datasheet. http://en.wikipedia.org/wiki/Standard_curve http://www.graphpad.com/curvefit/introduction5b.htm I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you very much for contacting us. The standard we have provided in the KIT is 2 ng/ml; which is lyophilized. We recommend adding 2 ml of Diluent Concentrate (10X) in the IL1 beta standard vial which will give final conc. of IL1 beta 1ng/ml. We also recommend preparing the standard curve using serial dilutions of standard. The information is nicely explained in protocol book attached on the datasheet. The standard curve positive slope will show the OD of standard corresponding to the concentration of IL1 beta standard. Now place the OD reading of your test sample on the standard curve and try to calculate the IL1 beta concentration from graph. If you have diluted samples six times then multiply the final value of IL1-beta by 6. Now coming to the substrate used p-Nitrophenyl Phosphate substrate is commonly used with Alkaline phosphatase enzyme; in the Kit we have used Streptavidin-Peroxidase conjugate which I guess is a HRP enzyme. To give you idea of what exactly the peroxiodase enzyme is, I have also sent an email to lab for confirmation of the enzyme; I will send you the information soon. I am however sure that the enzyme is not a phosphatase enzyme. Could you specify why p-Nitrophenyl Phosphate was used? Finally the chromogen substrate should be there in the Kit however in case of missing components we always send these as free of charge replacements to our customers. Could you confirm if you have contacted Abcam scientific support team regarding the missing components and discussed matter with them? I hope my suggestion will help to improve your results. Should you have any other question please do not hesitate to contact me. I will also look forward to receiving your reply soon.

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