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Mouse IL-6R alpha DuoSet ELISA

Catalog #: DY1830
12 Citations 1 Review Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY1830
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Mouse IL-6 R alpha ELISA Standard Curve
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Product Details
Procedure
Citations (12)
FAQs
Supplemental Products
Reviews (1)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Mouse IL-6R alpha DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
46.9 - 3,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse IL-6 sR. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

 

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A Mouse IL-6 R alpha ELISA Standard Curve View Larger

Mouse IL-6 R alpha ELISA Standard Curve

Product Datasheets

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Product Datasheet
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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: IL-6R alpha

IL-6 R alpha is a transmembrane subunit of the receptor for Interleukin-6. It associates with gp130 which is a shared component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 R alpha are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R alpha elicit responses from gp130-expressing cells that lack cell surface IL-6 R alpha. IL-6 plays important roles in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. IL-6, along with TNF-alpha and IL-1, drives the acute inflammatory response and the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. When dysregulated, it contributes to chronic inflammation in obesity, insulin resistance, inflammatory bowel disease, arthritis, sepsis, and atherosclerosis.

Long Name:
Interleukin 6 Receptor alpha
Entrez Gene IDs:
3570 (Human); 16194 (Mouse); 24499 (Rat); 102116430 (Cynomolgus Monkey)
Alternate Names:
CD126; gp80; IL-6 R alpha; IL6Q; IL6R alpha; IL-6R alpha; IL6R; IL-6R-1; IL6RA; IL-6Ra; IL6RQ; interleukin 6 receptor

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

 

Citations for Mouse IL-6R alpha DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

12 Citations: Showing 1 - 10
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  1. Cathepsin S provokes interleukin-6 (IL-6) trans-signaling through cleavage of the IL-6 receptor in vitro
    Authors: CM Flynn, Y Garbers, S Düsterhöft, R Wichert, J Lokau, CHK Lehmann, D Dudziak, B Schröder, C Becker-Pau, S Rose-John, S Aparicio-S, C Garbers
    Scientific Reports, 2020-12-10;10(1):21612.
    Species: Mouse
    Sample Types: Serum
  2. Anti-interleukin-6 receptor antibody treatment ameliorates postoperative adhesion formation
    Authors: N Uyama, H Tsutsui, S Wu, K Yasuda, E Hatano, XY Qin, S Kojima, J Fujimoto
    Sci Rep, 2019-11-26;9(1):17558.
    Species: Mouse
    Sample Types: Serum
  3. IL-6 exhibits both cis and trans signaling in osteocytes and osteoblasts, but only�trans�signaling promotes bone formation and osteoclastogenesis
    Authors: NE McGregor, M Murat, J Elango, IJ Poulton, EC Walker, B Crimeen-Ir, PWM Ho, JH Gooi, TJ Martin, NA Sims
    J. Biol. Chem., 2019-03-28;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. miR-146a modulates autoreactive Th17 cell differentiation and regulates organ-specific autoimmunity
    Authors: B Li, X Wang, IY Choi, YC Wang, S Liu, AT Pham, H Moon, DJ Smith, DS Rao, MP Boldin, L Yang
    J. Clin. Invest., 2017-09-05;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Intravitreal injection of anti-Interleukin (IL)-6 antibody attenuates experimental autoimmune uveitis in mice
    Authors: J Tode, E Richert, S Koinzer, A Klettner, U Pickhinke, C Garbers, S Rose-John, B Nölle, J Roider
    Cytokine, 2017-03-06;96(0):8-15.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. The soluble interleukin-6 receptor is a mediator of hematopoietic and skeletal actions of parathyroid hormone.
    Authors: Cho S, Pirih F, Koh A, Michalski M, Eber M, Ritchie K, Sinder B, Oh S, Al-Dujaili S, Lee J, Kozloff K, Danciu T, Wronski T, McCauley L
    J Biol Chem, 2013-01-07;288(10):6814-25.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. Cutting edge: Inhibition of IL-6 trans-signaling protects from malaria-induced lethality in mice.
    Authors: Wunderlich CM, Delic D, Behnke K, Meryk A, Strohle P, Chaurasia B, Al-Quraishy S, Wunderlich F, Bruning JC, Wunderlich FT
    J. Immunol., 2012-03-30;188(9):4141-4.
    Species: Mouse
    Sample Types: Serum
  8. Leukocyte ADAM17 Regulates Acute Pulmonary Inflammation.
    Authors: Arndt PG, Strahan B, Wang Y, Long C, Horiuchi K, Walcheck B
    PLoS ONE, 2011-05-16;6(5):e19938.
    Species: Mouse
    Sample Types: BALF
  9. Loss of CD4+ T cell IL-6R expression during inflammation underlines a role for IL-6 trans signaling in the local maintenance of Th17 cells.
    Authors: Jones GW, McLoughlin RM, Hammond VJ, Parker CR, Williams JD, Malhotra R, Scheller J, Williams AS, Rose-John S, Topley N, Jones SA
    J. Immunol., 2010-01-18;184(4):2130-9.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Essential roles of IL-6 trans-signaling in colonic epithelial cells, induced by the IL-6/soluble-IL-6 receptor derived from lamina propria macrophages, on the development of colitis-associated premalignant cancer in a murine model.
    Authors: Matsumoto S, Hara T, Mitsuyama K, Yamamoto M, Tsuruta O, Sata M, Scheller J, Rose-John S, Kado S, Takada T
    J. Immunol., 2009-12-30;184(3):1543-51.
    Species: Mouse
    Sample Types: Tissue Homogenates
  11. Direct B cell stimulation by dendritic cells in a mouse model of lupus.
    Authors: Wan S, Zhou Z, Duan B, Morel L
    Arthritis Rheum., 2008-06-01;8(0):1741-50.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  12. Classic interleukin-6 receptor signaling and interleukin-6 trans-signaling differentially control angiotensin II-dependent hypertension, cardiac signal transducer and activator of transcription-3 activation, and vascular hypertrophy in vivo.
    Authors: Coles B, Fielding CA, Rose-John S, Scheller J, Jones SA, O'Donnell VB
    Am. J. Pathol., 2007-07-01;171(1):315-25.
    Species: Mouse
    Sample Types: Plasma

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Mouse IL-6R alpha DuoSet ELISA
By Anonymous on 11/13/2019
Sample Tested: Serum