製品の概要

  • 製品名
    Mouse IL-1 beta ELISA Kit
    IL-1 beta キット 製品一覧
  • 検出方法
    Colorimetric
  • サンプルの種類
    Cell culture supernatant, Plasma
  • アッセイタイプ
    Sandwich (quantitative)
  • 検出感度
    < 5 pg/ml
  • 検出範囲
    2.74 pg/ml - 2000 pg/ml
  • 添加回収試験

    99 %

    特定サンプルでの回収試験
    サンプルの種類 平均 % 測定範囲
    Cell culture supernatant 98.64 89% - 110%
    Plasma 99.24 90% - 109%

  • ステップ
    Multiple steps standard assay
  • 種交差性
    交差種: Mouse
  • 製品の概要

    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.

  • アプリケーション
    適用あり: Sandwich ELISAmore details
  • 試験プラットフォーム
    Microplate

製品の特性

  • 保存方法
    Store at -20°C. Please refer to protocols.
  • 内容 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
  • 研究分野
  • 機能
    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.
  • 組織特異性
    Expressed in activated monocytes/macrophages (at protein level).
  • 配列類似性
    Belongs to the IL-1 family.
  • 翻訳後修飾
    Activation of the IL1B precursor involves a CASP1-catalyzed proteolytic cleavage. Processing and secretion are temporarily associated.
  • 細胞内局在
    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
  • 別名
    • 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
  • 参照データベース

関連製品

アプリケーション

Our Abpromise guarantee covers the use of ab100704 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Sandwich ELISA Use at an assay dependent concentration.

画像

  • Standard curve of msIL-1b in different diluents with background signal subtracted (duplicates; +/- SD).
  • IL-1b measured in mouse tissue lysates (expressed as per mg of extracted protein), with background signal subtracted (duplicates; +/- SD).
  • 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).
  • Representative standard curve using ab100704

  • Representative standard curve using ab100704

プロトコール

参考文献

This product has been referenced in:
  • Yang Z  et al. miR-143-3p regulates cell proliferation and apoptosis by targeting IGF1R and IGFBP5 and regulating the Ras/p38 MAPK signaling pathway in rheumatoid arthritis. Exp Ther Med 15:3781-3790 (2018). Read more (PubMed: 29581736) »
  • Xu X  et al. MFG-E8 inhibits Aß-induced microglial production of cathelicidin-related antimicrobial peptide: A suitable target against Alzheimer's disease. Cell Immunol N/A:N/A (2018). ELISA ; Mouse . Read more (PubMed: 29861070) »
See all 16 Publications for this product

レビューと Q&A

1-10 of 10 Abreviews or Q&A

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.
.

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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.

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Question
Answer

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

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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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Question
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.

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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.

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Question
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

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Question
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.

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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.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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