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    rat-il-6-elisa-kit-ab100772.pdf

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Immunology Innate Immunity Macrophage / Inflamm.
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Rat IL-6 ELISA Kit (ab100772)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2)Q&A (14)References (35)

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Representative Standard Curve

    Key features and details

    • Sensitivity: 30 pg/ml
    • Range: 40.96 pg/ml - 10000 pg/ml
    • Sample type: Cell culture supernatant, Plasma
    • Detection method: Colorimetric
    • Assay type: Sandwich (quantitative)
    • Reacts with: Rat

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    Overview

    • Product name

      Rat IL-6 ELISA Kit
      See all IL-6 kits
    • Detection method

      Colorimetric
    • Sample type

      Cell culture supernatant, Plasma
    • Assay type

      Sandwich (quantitative)
    • Sensitivity

      < 30 pg/ml
    • Range

      40.96 pg/ml - 10000 pg/ml
    • Recovery

      119 %

      Sample specific recovery
      Sample type Average % Range
      Cell culture supernatant 130.2 118% - 138%
      Plasma 122.8 109% - 138%
    • Assay duration

      Multiple steps standard assay
    • Species reactivity

      Reacts with: Rat
    • Product overview

      Abcam’s IL-6 Rat ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of rat IL-6 in plasma or cell culture supernatants.


      This assay employs an antibody specific for Rat IL-6 coated on a 96- well plate. Standards and samples are pipetted into the wells and IL-6 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Rat IL-6 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-6 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.


      Due to the relatively low abundance of IL-6 in normal rat plasma, this kit may not be suitable for use with this sample type. We have not been able to detect endogenous Rat IL-6 in normal serum with ab100772, only in serum spiked with Rat IL-6.

    • Platform

      Microplate

    Properties

    • Storage instructions

      Store at -20°C. Please refer to protocols.
    • Components 1 x 96 tests
      20X Wash Buffer Concentrate 1 x 25ml
      400X HRP-Streptavidin Concentrate 1 x 200µl
      5X Assay Diluent B 1 x 15ml
      Assay Diluent C 1 x 30ml
      Biotinylated anti-Rat IL-6 (lyophilized) 2 vials
      IL-6 Microplate (12 x 8 wells) 1 unit
      Recombinant Rat IL-6 Standard (lyophilized) 2 vials
      Stop Solution 1 x 8ml
      TMB One-Step Substrate Reagent 1 x 12ml
    • Research areas

      • Immunology
      • Innate Immunity
      • Macrophage / Inflamm.
      • Immunology
      • Innate Immunity
      • Cytokines
      • Interleukins
      • Neuroscience
      • Neurology process
      • Metabolism
      • Cancer
      • Invasion/microenvironment
      • Angiogenesis
      • Angiogenic growth factors
      • Cancer
      • Tumor immunology
      • Cytokines
      • Interleukins
      • Kits/ Lysates/ Other
      • Kits
      • ELISA Kits
      • ELISA Kits
      • Cytokines and cytokine receptors ELISA kits
      • Metabolism
      • Types of disease
      • Obesity
      • Metabolism
      • Types of disease
      • Metabolic disorders
    • Function

      Cytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Plays an essential role in the final differentiation of B-cells into Ig-secreting cells Involved in lymphocyte and monocyte differentiation. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation Acts on B-cells, T-cells, hepatocytes, hematopoeitic progenitor cells and cells of the CNS. Also acts as a myokine. It is discharged into the bloodstream after muscle contraction and acts to increase the breakdown of fats and to improve insulin resistance.
    • Involvement in disease

      Genetic variations in IL6 are associated with susceptibility to rheumatoid arthritis systemic juvenile (RASJ) [MIM:604302]. An inflammatory articular disorder with systemic-onset beginning before the age of 16. It represents a subgroup of juvenile arthritis associated with severe extraarticular features and occasionally fatal complications. During active phases of the disorder, patients display a typical daily spiking fever, an evanescent macular rash, lymphadenopathy, hepatosplenomegaly, serositis, myalgia and arthritis.
      Note=A IL6 promoter polymorphism is associated with a lifetime risk of development of Kaposi sarcoma in HIV-infected men.
    • Sequence similarities

      Belongs to the IL-6 superfamily.
    • Post-translational
      modifications

      N- and O-glycosylated.
    • Cellular localization

      Secreted.
    • Target information above from: UniProt accession P05231 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Alternative names

      • Interleukin BSF 2
      • B cell differentiation factor
      • B cell stimulatory factor 2
      • B-cell stimulatory factor 2
      • BSF 2
      • BSF-2
      • BSF2
      • CDF
      • CTL differentiation factor
      • Cytotoxic T cell differentiation factor
      • Hepatocyte stimulating factor
      • Hepatocyte stimulatory factor
      • HGF
      • HSF
      • Hybridoma growth factor
      • Hybridoma growth factor Interferon beta-2
      • Hybridoma plasmacytoma growth factor
      • IFN-beta-2
      • IFNB2
      • IL 6
      • IL-6
      • IL6
      • IL6_HUMAN
      • Interferon beta 2
      • Interferon beta-2
      • Interleukin 6
      • Interleukin 6 (interferon beta 2)
      • Interleukin BSF 2
      • Interleukin-6
      see all
    • Database links

      • Entrez Gene: 24498 Rat
      • SwissProt: P20607 Rat
      • Unigene: 9873 Rat

      Associated products

        Images

        • Representative Standard Curve
          Representative Standard Curve

          Representative Standard Curve using ab100772

        Protocols

        • Protocol Booklet

        Click here to view the general protocols

        Datasheets and documents

        • Datasheet
        • SDS
      • References (35)

        Publishing research using ab100772? Please let us know so that we can cite the reference in this datasheet.

        ab100772 has been referenced in 35 publications.

        • Alkhedaide AQ Anti-inflammatory Effect of Juniperus Procera Extract in Rats Exposed to Streptozotocin Toxicity. Antiinflamm Antiallergy Agents Med Chem 18:71-79 (2019). PubMed: 30474537
        • Sha H  et al. Rheinic acid ameliorates radiation-induced acute enteritis in rats through PPAR-?/NF-?B. Genes Genomics N/A:N/A (2019). PubMed: 31037524
        • Sun L  et al. The involvement of spinal annexin A10/NF-?B/MMP-9 pathway in the development of neuropathic pain in rats. BMC Neurosci 20:28 (2019). PubMed: 31208343
        • Gundala NKV & Das UN Arachidonic acid-rich ARASCO oil has anti-inflammatory and antidiabetic actions against streptozotocin?+?high fat diet induced diabetes mellitus in Wistar rats. Nutrition 66:203-218 (2019). PubMed: 31310962
        • Zhou R  et al. Recombinant CC16 inhibits NLRP3/caspase-1-induced pyroptosis through p38 MAPK and ERK signaling pathways in the brain of a neonatal rat model with sepsis. J Neuroinflammation 16:239 (2019). PubMed: 31775794
        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 16 Abreviews or Q&A

        AbTrial test for rat lung tissue lysate samples

        Inconclusive Excellent 5/5 (Ease of Use)
        Abreviews
        Abreviews
        Tried rat lung lysate (in PBS) on kit ab100772, all the readings are either negative or below detection range. So this kit is not a good fit for tissue lysate

        Abcam user community

        Verified customer

        Submitted Feb 09 2016

        assay of rat serum cytokine expression in adjuvant-induced arthritis

        Inconclusive Good 4/5 (Ease of Use)
        Abreviews
        Abreviews
        the product worked effectively.

        Newman Osafo

        Verified customer

        Submitted Dec 17 2014

        Question

        Can serum samples be run "neat" (without dilution) in the IL-4 Elisa - ab100770?

        Read More

        Abcam community

        Verified customer

        Asked on May 16 2013

        Answer


        The lab adviced the following:

        While serum and plasma samples maybe run undiluted, we do not usually recommend doing so. The reason is not because Assay Diluent A needs to be present in every sample but that testing undiluted serum/plasma samples increases the risk of experiencing matrix effects.


        Matrix effects occur when some component of the sample affects immunoreactivity of the target molecule, possibly by sequestering binding sites. Auto-antibodies, binding proteins, albumin, etc. may be possible culprits, but it’s hard to say for sure. The result is that the sample may read falsely high or low in the ELISA test, or may give a non-linear dilution response. Sometimes certain disease states contribute to matrix effects by altering the properties of the blood (highly lipemic samples, for instance); hemolysis may also cause similar problems. Matrix effects can be hard to predict and challenging to overcome.


        For this reason, we rarely recommend running plasma or serum samples undiluted, as matrix effects are more likely to occur. The lowest dilution factor we normally recommend starting with is 2-fold, even for the low-abundance cytokines. If the customer is concerned that their sample levels will be below detection, keep in mind the optional overnight standard/sample incubation can be performed to try and maximize the absorbances. If that doesn’t work, the customer may want to consider concentrating their samples or switching sample types (for example plasma) to see if that helps.

        Read More

        Abcam Scientific Support

        Answered on May 16 2013

        Question

        What is the normal amount of rat IL4 in serum?
        What is the normal amount of rat IL6 in serum?
        What is the recommended dilution range?

        Read More

        Abcam community

        Verified customer

        Asked on May 13 2013

        Answer


        I have the following information for you regarding the kits we discussed:


        ab46073:
        We have no data regarding normal values observed with IL-6 and IL-4 in rat but as free circulating cytokines are rather dangerous for living animal, the normal level is probably below the detection level.


        ab119548:
        Unfortunately, we have not tested the rat IL-6 ELISA on normal level in rat samples. We have only tested this ELISA using samples spiked with recombinant protein.


        ab100770 and ab100772:
        1) What is the normal amount of rat IL4 in serum?
        We tested the normal amount of IL-4 in healthy rat serum/plasma samples to be undetectable – 2 pg/ml.

        2) What is the normal amount of rat IL6 in serum?
        We tested the normal amount of IL-6 in healthy rat serum/plasma samples to be below the detectable limits of this ELISA. Thus, this kit may be more suitable for diseased/treated samples; however, there are some attached tips for increasing sensitivity the customer may use - see below.

        3) What is the recommended dilution range for rat serum for IL4 and IL6?
        We recommend a 2-fold dilution for serum/plasma samples for use with our IL-4 and IL-6 Rat ELISA kits.


        Some advice for what to do if samples are reading below detection limit and how the sensitivity of the kit can be increased:


        Incubate sample overnight at 4 degrees C
        Increase amount of biotinylated ab (by 1.5 fold or so – too much may increase background)
        Incubate biotinylated ab overnight at 4 degrees C
        Increase amount of HRP-strep (by about 1.5 fold or so – too much may increase background)
        Concentrate sample (using a spin column)



        Please note: it’s best to try just one of these modifications at a time. Implementing too many of these changes at once may cause high background.

        Read More

        Abcam Scientific Support

        Answered on May 13 2013

        Question

        Standard curve worked fine but samples (undiluted plasma) came out lower than blank.

        Read More

        Abcam community

        Verified customer

        Asked on Dec 21 2012

        Answer

        Thank you for your inquiry.

        I heard back from the lab that you should try diluting the plasma samples because this minimizes the chances of matrix effects which can lead to inaccurate detection or non-linear responses. Matrix effects happen when some component of the sample affects immunoreactivity of the target molecule, possibly by sequestering binding sites. Auto-antibodies, binding proteins, albumin, etc. may be possible culprits, but it can of course vary depending on the sample. Sometimes certain disease states contribute to matrix effects by altering the properties of the blood (highly lipemic samples, for instance); hemolysis may also cause similar problems. For this reason, we rarely recommend running plasma or serum samples undiluted, as matrix effects are more likely to occur. The quickest and easiest way to deal with matrix effects is to dilute the sample until you obtain a linear dilution response as this will dilute out the interfering components of the sample. Our recommended dilution for rat plasma samples for this kit is 2X, of course, optimal dilution ranges are always determined by the experimenter empirically.

        There may be other steps you can take to increase your detection:

        Incubate sample overnight at 4 degrees C
        Incubate biotinylated ab overnight at 4 degrees C
        Increase amount of HRP-strep (by about 1.5 fold or so – too much may increase background)




        Please note: it’s best to try just one of these modifications at a time. Implementing too many of these changes at once may cause high background.

        I hope this information helps. Please contact us with any other questions.

        Read More

        Abcam Scientific Support

        Answered on Dec 21 2012

        Question

        Apreciada
        Te adjunto más abajo en color rojo las respuestas a vuestros comentarios.
        Un saludo
        - Ambos kits se pidieron en Junio, si no se han utilizado hasta ahora es posible que haya habido degradación de los reactivos del mismo si no se ha almacenado a la temperatura adecuada. ¿Puedes confirmarme como se ha almacenado el kit, y el vial de estándar después de su reconstitución? ¿Estaba abierta la placa durante tiempo antes de utilizarse?
        -Los kits se almacenaron a su llegada en la cámara fría a 4ºC. Las cajas no se desprecintaron hasta el día en que se realizó el kit. El estándar se reconstituyó en el momento de realizar el kit, por lo que no fue necesario almacenarlo. La placa estaba cerrada, ya que como he comentado anteriormente, el kit no se desprecintó hasta el momento de realizar el ensayo.
        - Por el protocolo enviado entiendo que las muestras de suero no se diluyeron. Nosotros recomendamos una dilución mínima de 1/2 para muestras de suero o plasma. Si no se diluyen las muestras existe el riesgo de observar efecto matriz (este efecto ocurre cuando alguno de los compuestos de la muestra afectan a la immuno reactividad de la molécula target, enmascarando los sitios de unión). Algunos posibles causantes de ello son los anticuerpos propios, proteínas de unión, albumina etc… El resultado es que la muestra puede dar una lectura falsa, por encima o por debajo de los valores adecuados en un test ELISA, o incluso dar una respuesta no lineal.
        Incluso en algunos casos algunas enfermedades contribuyen a crear el efecto matriz alterando las propiedades de la sangre. Este efecto es difícil de predecir y de solucionar.
        Por ello no solemos recomendar usar muestras de plasma o suero sin diluir, para evitar este efecto. El factor de dilución mínimo recomendado es 1/2, incluso para la detección de citoquinas poco abundantes como IL1 beta e IL6.
        -A pesar de que en los datos que os envié no se observa, las muestras de suero si estaban diluidas a 1/3.
        -También sugeriría incubar el anticuerpo por más tiempo. Al menos 2.5 horas, pero lo ideal es dejarlo actuar durante la noche para incrementar la señal.
        -Las muestras fueron incubadas durante toda la noche a 4ºC en agitación suave.
        Si te parece y estás de acuerdo te propongo ensayar los cambios sugeridos y observar si la señal aumenta. En caso contrario, ya que ambos kits están cubiertos por la garantía Abpromise, se te reemplazarían los kits o reembolsaría el importe de los mismos.
        -Debido a que se trataba de muestras de suero de ratas, disponíamos de muy poca muestra, por lo que actualmente nos es imposible volver a analizar las muestras. Por esta razón propongo que nos rembolséis el importe de los dos kits. En un futuro, cuando volvamos a generar muestras, adquiriremos nuevos kits.

        Read More

        Abcam community

        Verified customer

        Asked on Dec 07 2012

        Answer

        Perdona por la espera.
        Es muy posible que el motivo de la baja absorbancia observada en las placas se deba a la perdida de actividad de alguno de los componentes del kit, debido a su degradación por haberse mantenido durante tantos meses a 4C.
        Siempre recomendamos adquirir los productos, especialmente los kits de actividad, cuando vayan a utilizarse. Si se almacenan durante más de un mes es aconsejable mantenerlos a -20C para evitar posibles pérdidas de actividad.
        La señal del estándar para el kit de IL6 es baja, sin embargo para el kit IL1beta los valores son aceptables. En este último caso el problema es quizás el background, que provoca la perdida de sensibilidad de la recta del estándar. Adjunto un par de documentos con algunos consejos para obtener señales intensas del estándar en las placas y para disminuir el grado de background.
        De todas formas, el problema más común en experimentos de determinación de IL6 e IL1beta, no tiene que ver con el ensayo en sí, sino simplemente con los bajos valores de estas citoquinas.
        En suero y plasma estas se encuentran en niveles bajos normalmente. Sin embargo, hay ciertos parámetros que deben mantenerse en estos ensayos para maximizar la probabilidad de que las muestras se encuentren dentro del rango detectable del ensayo:
        - Incubar muestras y estándar durante la noche a 4C.
        - Incrementar la cantidad de anticuerpo de detección (en 1,5. Demasiado incremento provocaría background)
        -Incrementar la cantidad de HRP Streptavidina (de nuevo en 1,5 para evitar la aparición de background)
        - Concentrar las muestras (usando, por ejemplo, una columna).
        Se recomienda usar cada una de estas modificaciones por separado para evitar de nuevo la aparición de background.
        De todas formas dado que ambos kits se encuentran en garantía si deseáis recibir un reembolso o reemplazo no dudes en comunicármelo.
        Espero que esta información sea útil. Espero tu respuesta al respecto.

        Read More

        Abcam Scientific Support

        Answered on Dec 07 2012

        Question

        Hola
        Este mail si lo he recibido perfectamente.
        Saludos

        Read More

        Abcam community

        Verified customer

        Asked on Dec 05 2012

        Answer

        Gracias por mantenerte a la espera.
        Después de discutir el caso con mis compañeros, hemos comentado lo siguiente:
        - Ambos kits se pidieron en Junio, si no se han utilizado hasta ahora es posible que haya habido degradación de los reactivos del mismo si no se ha almacenado a la temperatura adecuada. ¿Puedes confirmarme como se ha almacenado el kit, y el vial de estándar después de su reconstitución? ¿Estaba abierta la placa durante tiempo antes de utilizarse?
        - Por el protocolo enviado entiendo que las muestras de suero no se diluyeron. Nosotros recomendamos una dilución mínima de 1/2 para muestras de suero o plasma. Si no se diluyen las muestras existe el riesgo de observar efecto matriz (este efecto ocurre cuando alguno de los compuestos de la muestra afectan a la immuno reactividad de la molécula target, enmascarando los sitios de unión). Algunos posibles causantes de ello son los anticuerpos propios, proteínas de unión, albumina etc…
        El resultado es que la muestra puede dar una lectura falsa, por encima o por debajo de los valores adecuados en un test ELISA, o incluso dar una respuesta no lineal.
        Incluso en algunos casos algunas enfermedades contribuyen a crear el efecto matriz alterando las propiedades de la sangre. Este efecto es difícil de predecir y de solucionar.
        Por ello no solemos recomendar usar muestras de plasma o suero sin diluir, para evitar este efecto. El factor de dilución mínimo recomendado es 1/2, incluso para la detección de citoquinas poco abundantes como IL1 beta e IL6.
        -También sugeriría incubar el anticuerpo por más tiempo. Al menos 2.5 horas, pero lo ideal es dejarlo actuar durante la noche para incrementar la señal.
        Si te parece y estás de acuerdo te propongo ensayar los cambios sugeridos y observar si la señal aumenta. En caso contrario, ya que ambos kits están cubiertos por la garantía Abpromise, se te reemplazarían los kits o reembolsaría el importe de los mismos.
        Espero que esta información sea útil. No dudes en contactarme para cualquier comentario que puedas tener al respecto.

        Read More

        Abcam Scientific Support

        Answered on Dec 05 2012

        Question

        Apreciada
        Adjunto te envío los cuestionarios, tal y como dijiste en tu último e-mail.
        Atentamente

        Read More

        Abcam community

        Verified customer

        Asked on Nov 19 2012

        Answer

        Gracias por mandarme los cuestionarios rellenos.

        ¿Podrías por favor confirmarme como se llevo a cabo la preparación de la solución de HRP Streptavidina?

        Recientemente se han cambiado los protocolos de algunos de nuestros kits, y antes la solución de Streptavidina-HRP estaba diluida 1000x, mientras que en la nueva versión se ha concentrado a 200X.

        Si se hubiera llevado a cabo una preparación de la solución teniendo en cuenta la dilución 1/1000 esto explicaría por qué los resultados de absorbancia son tan bajos.

        Si no fuera esta la causa, trataremos de encontrar otra solución en la mayor brevedad posible.

        Read More

        Abcam Scientific Support

        Answered on Nov 19 2012

        Question

        Apreciados Sres.,
        Recientemente les compramos los kit de ELISA IL-6 y IL-1β en rata (junto a otro grupo de kits para detectar otras citoquinas) a mediados de Junio.
        Al realizar los kits de Elisa hemos obtenido una absorbancia muy baja en la curva estándar, por supuesto, realizando el ELISA siguiendo el procedimiento descrito en el kit y sin ningún tipo de desviación.

        Debido a que en mayo de este mismo año nos encontramos con una situación similar con un kit de ELISA IL-6 (adjunto e-mail mas abajo), realizamos la prueba de la dilución de la Streptavidina, como nos indicasteis la última vez, y no se ha observado coloración azul.

        Para nosotros es muy importante poder realizar los ensayos correctamente en la mayor brevedad posible, por lo que, considerando que el problema reside al parecer en alguno de los componentes del kit, agradecería nos remplazaran los productos por otros en buenas condiciones

        Muchísimas gracias de antemano.
        Reciba un cordial saludo

        Read More

        Abcam community

        Verified customer

        Asked on Nov 12 2012

        Answer

        Gracias por contactarnos.

        Vuelvo a pediros disculpas por las molestias causadas, y siento mucho que estos kits no hayan dado el resultado esperado.

        Para poder investigar este caso, evitar que vuelva a repetirse, y desde luego estar seguros que los kits que os mandemos a modo de reemplazo están en perfectas condiciones, te agradecería que mandaras mas detalles de los resultados obtenidos.

        Te pediría por favor que me reenviaras el cuestionario adjunto relleno, con cuantos más detalles mejor acerca del almacenamiento del kit, y cualquier observación que hayáis podido hacer que os haya llamado la atención al realizar el ensayo (como cambios de color).

        Te agradezco también que me envíes los valores obtenidos para poder comentarlos con mis compañeros del laboratorio, y así analizar la procedencia del fallo del kit.

        Agradezco mucho tu comprensión, y confío en que este problema no vuelva a repetirse en el futuro.

        Read More

        Abcam Scientific Support

        Answered on Nov 12 2012

        Question

        Inquiry: Dear Sir/Madam, We are a group of researchers working in the department of pharmacology at Sahlgrenska Academy, University of Gothenburg, Sweden. We are interested in one of the products that your company offers (the kit https://www.abcam.com/IL6-Rat-ELISA-Kit-1-x-96-Well-Plate-ab100772.html ). It would be appreciated if you could tell us the cost of the kit and the delivery time. Yours faithfully,

        Read More

        Abcam community

        Verified customer

        Asked on Sep 27 2012

        Answer

        Thank you for your enquiry and your interest.

        This kit is in stock and if you place the order today, we could deliver it by Friday the 28th September. The unit price is €490.00. I would advise you to read the datasheet as well as the on-line Protocol Booklet carefully to make sure that this productsuitsyour need.

        https://www.abcam.com/ps/products/100/ab100772/documents/ab100772-IL6-Rat-ELISA-Kit-ab100772-protocol-plain-v2.pdf

        Ifyou need any further assistance in the future, please do not hesitate to contact me.

        Read More

        Abcam Scientific Support

        Answered on Sep 27 2012

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