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Immunology Innate Immunity Cytokines Interleukins
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Mouse IL-1 beta ELISA Kit (ab100704)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (10)References (42)

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Sandwich ELISA - IL-1 beta Mouse ELISA Kit (ab100704)
  • Sandwich ELISA - IL-1 beta Mouse ELISA Kit (ab100704)
  • Sandwich ELISA - IL-1 beta Mouse ELISA Kit (ab100704)
  • Typical standard curve
  • Typical standard curve

Key features and details

  • Sensitivity: 5 pg/ml
  • Range: 2.74 pg/ml - 2000 pg/ml
  • Sample type: Cell culture supernatant, Plasma
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Mouse

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Overview

  • Product name

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

    Colorimetric
  • Sample type

    Cell culture supernatant, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 5 pg/ml
  • Range

    2.74 pg/ml - 2000 pg/ml
  • Recovery

    99 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 98.64 89% - 110%
    Plasma 99.24 90% - 109%
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Abcam’s IL-1 beta Mouse ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse IL-1 beta in plasma and cell culture supernatants.


    This assay employs an antibody specific for mouse IL-1 beta coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-1 beta present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse IL-1 beta antibody is added. After washing away unbound biotinylated antibody, HRP conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IL-1 beta bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


    We have not been able to detect the endogenous Mouse IL-1 beta in normal serum with ab100704, only in serum spiked with Mouse IL-1 beta.


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

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    200X HRP-Streptavidin Concentrate 1 x 200µl
    20X Wash Buffer Concentrate 1 x 25ml
    5X Assay Diluent B 1 x 15ml
    Assay Diluent A 1 x 30ml
    Biotinylated anti-mouse IL-1 beta 2 vials
    IL-1 beta Microplate (12 x 8 wells) 1 unit
    Recombinant Mouse IL-1 beta Standard (lyophilized) 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 1 x 12ml
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cytokines and cytokine receptors ELISA kits
    • Metabolism
    • Types of disease
    • Obesity
  • 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: 16176 Mouse
    • SwissProt: P10749 Mouse
    • Unigene: 222830 Mouse

    Associated products

    • SimpleStep ELISA kits

      • Mouse IL-1 beta ELISA Kit (ab197742)

    Images

    • Sandwich ELISA - IL-1 beta Mouse ELISA Kit (ab100704)
      Sandwich ELISA - IL-1 beta Mouse ELISA Kit (ab100704)
      Standard curve of msIL-1b in different diluents with background signal subtracted (duplicates; +/- SD).
    • Sandwich ELISA - IL-1 beta Mouse ELISA Kit (ab100704)
      Sandwich ELISA - IL-1 beta Mouse ELISA Kit (ab100704)
      IL-1b measured in mouse tissue lysates (expressed as per mg of extracted protein), with background signal subtracted (duplicates; +/- SD).
    • Sandwich ELISA - IL-1 beta Mouse ELISA Kit (ab100704)
      Sandwich ELISA - IL-1 beta Mouse ELISA Kit (ab100704)
      IL-1b detected in supernatants from RAW 246.7 control cells (C) or cells stimulated for 24 hours with 50 ng x mL-1 of PMA (ab120297) (P), or with PMA and 1 ug x mL-1 of LPS (Sigma) (P+L) for the last 6 hours. Results shown after background signal was subtracted (duplicates +/- SD).
    • Typical standard curve
      Typical standard curve

      Representative standard curve using ab100704

    • Typical standard curve
      Typical standard curve

      Representative standard curve using ab100704

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (42)

    Publishing research using ab100704? Please let us know so that we can cite the reference in this datasheet.

    ab100704 has been referenced in 42 publications.

    • Ma Y  et al. Mangiferin Relieves Lipopolysaccharide-Induced Injury by Up-Regulating miR-181a via Targeting PTEN in ATDC5 Cells. Front Pharmacol 11:137 (2020). PubMed: 32210798
    • Wang J  et al. Knockdown of long non-coding RNA PVT1 induces apoptosis of fibroblast-like synoviocytes through modulating miR-543-dependent SCUBE2 in rheumatoid arthritis. J Orthop Surg Res 15:142 (2020). PubMed: 32293498
    • Zhao Q  et al. Chitoheptaose Promotes Heart Rehabilitation in a Rat Myocarditis Model by Improving Antioxidant, Anti-Inflammatory, and Antiapoptotic Properties. Oxid Med Cell Longev 2020:2394704 (2020). PubMed: 32351668
    • Piao X  et al. Picroside II Improves Severe Acute Pancreatitis-Induced Intestinal Barrier Injury by Inactivating Oxidative and Inflammatory TLR4-Dependent PI3K/AKT/NF-?B Signaling and Improving Gut Microbiota. Oxid Med Cell Longev 2020:3589497 (2020). PubMed: 32351672
    • Luo Q  et al. NEMO-binding domain peptides alleviate perihematomal inflammation injury after experimental intracerebral hemorrhage. Neuroscience 409:43-57 (2019). PubMed: 31047976
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 10 Abreviews or Q&A

    Question

    I am interested in measuring mouse IL-1 beta in samples of plasma and tissue, is it possible to use only one type of kit (ab100704 or ab100705) for both? Which is the difference between them?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 09 2015

    Answer



    It is possible to use the non-lysate kit ab100704 (IL-1 beta (Interleukin-1 beta) Mouse ELISA Kit) and use your own lysis buffers (non-denaturing, with a mild detergent like RIPA/NP40/Triton X). The cell lysate kit (ab100705) contains this buffer and a different assay diluent, but the different assay diluent is not required to run the assay correctly with tissue/cell lysate samples.

    I can recommend to use a detergent based on our sample preparation tips here:

    Avoid using >0.1% SDS or other strongly denaturing detergents. In general, non-ionic detergents such as Triton X-100 or NP-40 are best, although zwitterionic detergents such as CHAPS, or mild ionic detergents such as sodium deoxycholate will work.
    Use no more than 2% v/v total detergent
    Avoid the use of sodium azide
    Avoid using >10 mM reducing agents, such as dithiothreitol or mercaptoethanols





    We strongly recommend adding a protease inhibitor cocktail to the lysis buffer prior to homogenization. Since susceptibility to proteolytic cleavage and the type of proteases present in the lysate vary, we do not recommend a specific product. Instead, your choice of which combination of protease inhibitors to use should be based upon a literature search for your protein(s) of interest and/or tissue or cell type. Phosphatase inhibitors may be used but are not necessary unless the antibodies used in the kit specifically recognize phosphorylated forms of the protein.

    Choices of the method for lysis and homogenization include glass-bead “smash,” douncing, freezethaw, sonication and crushing frozen tissue with a mortar and pestle, or even a combination of these. There is no best method for all sample types; your choice of method should be made following a brief search of the literature to see how samples similar to yours have been prepared in previous investigations.

    After homogenization, centrifuge the lysates to remove cell/tissue debris (5 min @ 10,000 x g or 10 min @ 5,000 x g) and save the supernatant. Unless testing fresh, lysates should be frozen as soon as possible and stored at -20°C (or -80°C, if possible). Centrifuge them again before incubating with any immunoassay. Next, determine the protein concentration of your lysates using a total protein assay not inhibited by detergents (such as the Bicinchoninic acid (BCA) assay) and normalize the volume of each sample used to deliver the same amount of total protein for each assay. Note: The Bradford assay is not recommended as it can be inhibited by the presence of detergents.

    Since different cells and tissues may contain different amounts of protein, as starting point, we suggest using 500 µL of lysis buffer per 1x106 cells or 10 mg tissue. You may have to adjust this based upon your results. Your target total protein concentration of the homogenate should be at least 1,000 µg/mL, but 2,000 µg/mL or more would be better.
    .

    Read More

    Anja Hoffmann

    Abcam Scientific Support

    Answered on Sep 09 2015

    Question

    I am concerned that since I am using cell culture supernatant as my sample, soluble IL-1 beta receptors in the supernatant will compete with the antibodies in the kit to bind to IL-1 beta, making the amount of measured IL-1 beta lower than what it actually is in my sample. Are there any components/buffers in the kit that control for this? If I suspect low levels of IL-1 beta, is it ok to run the assay without diluting my samples with assay diluent B?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 26 2014

    Answer

    It is not clear if the kit is detecting IL-1 beta free or bound and we do not have any components or buffers that are specifically made to unbind IL-1 beta from the receptor.

    Please see below available antibody specificity information:
    Capture Antibody
    Source: Monoclonal Rat IgG1
    Specificity: Detects mouse IL­1β/IL­1F2
    Immunogen: E. coli­derived recombinant mouse IL­1β/IL­1F2 (aa 118-269)
    Accession # P10749

    Detection Antibody
    Source: Polyclonal Goat IgG
    Specificity: Less than 4% cross­reactivity with recombinant rat (rr) IL­1β is observed and less than 0.05% cross­reactivity with rhIL­1β and rmIL­1α is observed.
    Immunogen: E. coli­derived recombinant mouse IL­1β (aa 118-269)
    Accession # NP_032387

    As for running the samples undiluted, we would not recommend this due to matrix effects.

    Read More

    Kevin Hanson

    Abcam Scientific Support

    Answered on Feb 26 2014

    Question

    I also wonder that whether heparin or EDTA
    affect ELISA procedure.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 10 2014

    Answer

    I confirm this kit (ab100704) is compatible with heparin and EDTA.

    Read More

    Ariana Veiga

    Abcam Scientific Support

    Answered on Feb 10 2014

    Question

    Hola,
    Estoy buscando kits de ELISA para detección de citocinas (en particular, IL-1 beta, IL-17 e IFN gamma, pero si van bien, luego podrían ser más), pero necesitaría que detectaran citocinas tanto humanas como de ratón. Veo que ustedes tienen varios kits, pero todos reconocen la proteína humana o la de ratón. Ustedes tienen información sobre reacción cruzada? Es posible que alguno de estos kits reconozca las dos proteínas?
    Gracias!

    Read More

    Abcam community

    Verified customer

    Asked on Apr 12 2013

    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.

    Read More

    Abcam Scientific Support

    Answered on Apr 12 2013

    Question


    Thanks for your reply. The problem we have here is that we believe that the ELISA kit we have in at the moment (ebioscience kit) detects both mouse active and mouse pro IL-1b at different affinities. In more detail, we think that the active IL-1b is detected around 10X better than the pro-IL-1beta. Obviously this is not much good to our lab and so we need a new kit to measure both. It is however, it is not financially viable for me to order in a whole range of kits from different companies in order to find a kit that will detect btw forms of the cytokine equally. Instead I was hoping that you may be able to give me a sample of your kit. In return, we could provide some information on the specificity of the ELISA. As an additional incentive to offer a sample, we do a lot of IL-1b work in our lab and so, if the kit does detect both equally, we would most likely order many more kits in the future. I hope you understand our situation.
    Many thanks

    Read More

    Abcam community

    Verified customer

    Asked on Jan 07 2013

    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.

    Read More

    Abcam Scientific Support

    Answered on Jan 07 2013

    Question

    Buenos días,
    te adjunto el cuestionario que me habéis pedido rellenar.
    Un saludo,

    Read More

    Abcam community

    Verified customer

    Asked on Jan 03 2013

    Answer

    Te pido disculpas por la espera.

    He estado revisando los datos enviados con ayuda de mis compañeros del laboratorio. Los valores del estándar son débiles, pero no demasiado, alcanzando una densidad óptica de 0.875. Esta señal podría incrementarse algo, pero el problema principal son algunas muestras cerca del valor del ruido de fondo.

    Sin embargo la mayoría de los valores para las muestras están por encima del nivel del background, y sería posible extraer información útil de ellas. Si aun tenéis posibilidad de repetir el ensayo os sugiero reducir la dilución de las muestras, a 1:2, para incrementar la absorbancia de las muestras a la región media de la curva de estándares, que es la zona más linear y precisa.

    En principio los resultados son correctos. Simplemente, para incrementar la absorbancia es importante mantener el kit y sus componentes en las condiciones de almacenamiento recomendadas (a -20C si se mantiene durante largos periodos) y seguir las siguientes pautas:

    - Reducir la dilución de las muestras.

    - Cuando se prepare el estándar es importante centrifugar el vial antes de nada, y asegurarse de disolver todo el polvo de estándar al reconstituirlo. No usar el vortex para la reconstitución del estándar, puesto que desestabiliza la proteína. Una vez reconstituido debe usarse inmediatamente, o congelarse para usarse más adelante.

    - Las diluciones de estándar deben mantenerse en hielo durante su preparación, aunque el ensayo ELISA se realice en condiciones de temperatura ambiente.

    Espero que estas pautas consigan mejorar el resultado e incrementar la señal. Si no fuera así, o si tuvierais cualquier otra consulta, no dudes en contactarnos de nuevo.

    Read More

    Abcam Scientific Support

    Answered on Jan 03 2013

    Question

    I need to be able to detect both active and pro_IL-1beta from mouse cell culture supernatant and lysates.
    > I am looking for a way to quantitatively detect active and proIL-1beta separately. I have an ELISA that can pick up the pro form on its own but now need either an ELISA that will pick up both equally or just the active form. Do you have any products that will help me?
    > Thanks
    >
    >

    Read More

    Abcam community

    Verified customer

    Asked on Dec 19 2012

    Answer

    Thank you for your patience.

    I am still waiting on more information on ab108866.

    But I am happy to provide the following information about ab100704 and ab100705:

    The antibody pair used in the Mouse IL-1 beta Sandwich ELISA kits was raised against an immunogen which was the mature/cleaved form of the IL-1 beta protein consisting of amino acids 118-269. Therefore we know that the kit can detect the active/mature form.

    Whether the kit can also detect the propeptide however has not been determined so accordingly we don’t know if this kit can differentiate between the propeptide and the mature form. The only way to determine this would be to do either epitope mapping (which we haven’t done as it is time-consuming and costly) or to simply spike in only the propeptide and see if the kit detects it.

    Because the propeptide does contain the amino acid sequence of the mature peptide though, if it’s a linear epitope then likely the kit would also detect the propeptide. If the epitope is confirmational or some post-translational modification blocks the binding site, then the kit would likely not detect the propeptide). This is all speculation of course because we don’t know the epitope.

    I hope this information is helpful. Please do not hesitate to contact em again with any further questions.

    Read More

    Abcam Scientific Support

    Answered on Dec 19 2012

    Question

    protocol pour ab100704

    Read More

    Abcam community

    Verified customer

    Asked on Nov 28 2012

    Answer

    Merci de nous avoir contactés.

    Suite à notre conversation téléphonique, j’ai contacté le laboratoire. Ils m’ont répondu que diluent B fonctionne tout simplement mieux que le diluant A dans la préparation de l'anticorps de détection (c'est à dire qu'il produit un signal plus fort). Diluent A peut aussi être utilisé à l’étape 6 du protocole mais ne donnera pas des signaux aussi forts donc diluent B est recommandé même si vous utiliser des échantillons de sérum ou plasma.

    Par contre, pour la préparation de HRP-streptavidine, diluent A ne peut pas être utilisé. Ceci est parce que diluant A contient de l'azoture de sodium qui inhibe l'activité de HRP. Par conséquent diluant B devrait être utilisé pour préparer le HRP-streptavidine.

    J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

    Monoclonal de lapin gratuit pour tout achat d'anticorps primaire, dans la limite des stocks disponibles. Notez le code "RABMAB-XBSMG" sur votre prochaine commande d’anticorps primaire. Pour plus d'informations, visitez le lien suivant: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

    Abcam Scientific Support

    Answered on Nov 28 2012

    Question

    Does this kit detect the pro and mature forms of IL1 beta?

    Read More

    Abcam community

    Verified customer

    Asked on Aug 08 2012

    Answer

    Thank you very much for your call earlier this week and for your patience while I have been in touch with the lab.

    The mouse IL1 beta ELISA kit detects the mature form of IL-1beta. It has not been tested for detection of the pro form, so whether or not the antibodies detect both forms is unfortunately unknown.

    I am sorry that we do not have more information in this case. If you have any further questions or if there is anything else that we can do for you, please let me know and I'll be happy to help.

    Read More

    Abcam Scientific Support

    Answered on Aug 08 2012

    Question

    Bonjour,
    J'aimerai avoir plus de détails, sur l'anticorps IL-1 beta mouseutilisé dans le kit ELISA suivant: ab 100704.
    - Quelle est la référence de l'anticorps utilisé ?
    - Cet anticorps reconnait t-il spécifiquement l'IL-1 beta mature(c'est à dire la protéine IL-1 beta qui a été clivée par la caspase 1) ?
    Merci par avance.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 09 2012

    Answer

    Merci de nous avoir contactés.

    Le kit https://www.abcam.com/ab100704 est un kit d'ELISA sandwich, il contient donc 2 anticorps. L'anticorps de capture est un monoclonal de rat (numéro de clone non déterminé) et l'anticorps de détection est un polyclonal de chèvre.
    Ces 2 anticorps, spécifiques de l'IL-1 beta murine (P10749, http://www.uniprot.org/uniprot/P10749) ont été développés pour ce kit et ne sont malheureusement pas commercialisés séparément.

    J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

    Read More

    Abcam Scientific Support

    Answered on Jul 09 2012

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