Mouse IL-1 beta ELISA Kit (ab100704)

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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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.

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

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.

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.

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.

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.

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

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.

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.