Overview

  • Product name

    Mouse IL-6 R alpha ELISA Kit
    See all IL-6R kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Mouse serum 5 1.8%
    Inter-assay
    Sample n Mean SD CV%
    Mouse serum 3 12%
  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    12.6 pg/ml
  • Range

    31.3 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 85 80% - 89%
    Serum 105 103% - 106%
    Cell culture media 88 84% - 93%
    Heparin Plasma 103 101% - 105%
    EDTA Plasma 103 102% - 103%
    Citrate Plasma 104 100% - 107%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rat, Rabbit, Goat, Guinea pig, Hamster, Cow, Dog, Human, Pig
  • Product overview

    Abcam’s IL-6 R alpha (Interleukin-6 Receptor alpha) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-6 R alpha protein in mouse serum, plasma, urine, and cell culture supernatant.

    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Interleukin-6 Receptor (IL-6R, IL-6 Receptor), also known as Interleukin-6 Receptor subunit alpha and membrane glycoprotein 80, is part of a receptor that binds Interleukin-6 (IL-6). IL-6 Receptor weakly binds IL-6 with a low affinity, and then is presented to gp130 to facilitate transition into a high affinity signaling competent hexamer. Activation may lead to regulation of the immune response, acute-phase reactions and hematopoiesis. The production of IL-6 Receptor has been implicated in the pathogenesis of many diseases, including multiple myeloma, auto immune diseases and prostate cancer.IL-6 Receptor has two isoforms. The first is a transmembrane protein in which the N-terminal extracellular portion can be released by proteolytic cleavage. The second isoform is a secreted form that lacks the transmembrane and intracellular portions. This assay is directed at the extracellular portion of IL-6 Receptor and detects both the soluble and membrane bound forms.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse IL-6 R alpha Capture Antibody 1 x 600µl
    10X Mouse IL-6 R alpha Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BR 1 x 6ml
    Mouse IL-6 R alpha Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Part of the receptor for interleukin 6. Binds to IL6 with low affinity, but does not transduce a signal. Signal activation necessitate an association with IL6ST. Activation may lead to the regulation of the immune response, acute-phase reactions and hematopoiesis.
    Low concentration of a soluble form of IL6 receptor acts as an agonist of IL6 activity.
  • Tissue specificity

    Isoform 2 is expressed in peripheral blood mononuclear cells and weakly found in urine and serum.
  • Sequence similarities

    Belongs to the type I cytokine receptor family. Type 3 subfamily.
    Contains 1 fibronectin type-III domain.
    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
  • Domain

    The two fibronectin type-III-like domains, contained in the N-terminal part, form together a cytokine-binding domain.
    The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell-surface receptor binding.
  • Post-translational
    modifications

    A short soluble form may also be released from the membrane by proteolysis.
  • Cellular localization

    Secreted and Basolateral cell membrane.
  • Information by UniProt
  • Alternative names

    • CD126
    • CD126 antigen
    • Gp80
    • IL 6R alpha
    • IL-6 receptor subunit alpha
    • IL-6R 1
    • IL-6R subunit alpha
    • IL-6R-1
    • IL-6R-alpha
    • IL-6RA
    • Il6r
    • IL6R1
    • IL6RA
    • IL6RA_HUMAN
    • IL6Ralpha
    • Interleukin 6 receptor
    • Interleukin-6 receptor subunit alpha
    • Membrane glycoprotein 80
    • MGC104991
    • OTTHUMP00000034320
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab203360 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 concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Standard curve comparison between mouse IL-6 R alpha SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 5-fold increase in sensitivity.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of IL-6 R alpha were measured in duplicate and interpolated from the IL-6 R alpha standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-6 R alpha concentration was determined to be 20.9 ng/mL in mouse serum, 19.1 ng/mL in mouse plasma (Citrate), and 17.1 ng/mL in mouse plasma (EDTA).

  • The concentrations of IL-6 R alpha were measured in duplicate and interpolated from the IL-6 R alpha standard curve and corrected for sample dilution.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean IL-6 R alpha concentration was determined to be 17,650 pg/mL in mouse plasma (heparin), and 849 pg/mL in J774A.1 cell culture supernatant.

  • J774A.1 cells were grown in the absence (unstimulated) or presence of Phorbol Myristate Acetate (PMA) and phytohemagglutinin (PHA) (stimulated) for 3 days. IL-6 R alpha was measured in 2-fold diluted cell culture supernatants of unstimulated and PMA/PHA stimulated J774A.1 and cell culture media. Measured values were interpolated from the IL-6 R alpha Standard Curve diluted in Sample Diluent NS and corrected for dilution factor. Mean of duplicate values +/-SD are graphed, 568 pg/mL unstimulated, 849 pg/mL stimulated, and undetectable in media.

Protocols

References

This product has been referenced in:

  • Serizawa K  et al. Anti-IL-6 receptor antibody improves pain symptoms in mice with experimental autoimmune encephalomyelitis. J Neuroimmunol 319:71-79 (2018). ELISA ; Mouse . Read more (PubMed: 29685293) »
  • Lee EJ  et al. Radiation Inhibits Interleukin-12 Production via Inhibition of C-Rel through the Interleukin-6/ Signal Transducer and Activator of Transcription 3 Signaling Pathway in Dendritic Cells. PLoS One 11:e0146463 (2016). Read more (PubMed: 26745884) »
See all 2 Publications for this product

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