Mouse IL-27 p28/IL-30 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY1834
Ancillary Products Available
Mouse IL-27 p28 / IL-30 ELISA Standard Curve
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Product Details
Procedure
Citations (10)
FAQs
Supplemental Products
Reviews (2)

Mouse IL-27 p28/IL-30 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 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-27p28. 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

Mouse IL-27 p28 / IL-30 ELISA Standard Curve

Product Datasheets

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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-27 p28/IL-30

IL-27 is a heterodimeric cytokine that is composed of EBI3 (which is related to the p40 subunit of IL-12 and IL-23) and p28 (which is related to the p35 chain of IL-12). IL-27 is expressed by monocytes, endothelial cells, and dendritic cells, and it signals through a receptor complex composed of IL-27 R alpha/WSX-1/TCCR and gp130. IL-27 induces IL-12 receptor expression on naïve CD4+ T cells, sensitizing them to subsequent IL-12 induced Th1 development. It also functions as an anti-inflammatory cytokine by limiting T and NKT cell activity.

Long Name:
Interleukin 27 p28
Entrez Gene IDs:
246778 (Human); 246779 (Mouse)
Alternate Names:
IL27 p28; IL-27 p28; IL-30

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-27 p28/IL-30 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.

10 Citations: Showing 1 - 10
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  1. TLR induced IL-27 plays host-protective role against B16BL6 melanoma in C57BL/6 mice
    Authors: A Patidar, S Selvaraj, M Chakravart, I Guha, A Bhuniya, S Bera, S Dhar, K Roy, R Baral, D Chattopadh, C Pal, B Saha
    Cytokine, 2022;154(0):155871.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Staphylococcus aureus-induced immunosuppression mediated by IL-10 and IL-27 facilitates nasal colonisation
    Authors: AM Kelly, JM Leech, SL Doyle, RM McLoughlin
    PloS Pathogens, 2022;18(7):e1010647.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  3. Interleukin-30 Suppresses Not Only CD4+ T Cells but Also Regulatory T Cells in Murine Primary Biliary Cholangitis
    Authors: HW Chen, CI Lin, YH Chuang
    Biomedicines, 2021;9(8):.
    Species: Mouse
    Sample Types: Serum
  4. The Role of Alveolar Macrophages in the Improved Protection against Respiratory Syncytial Virus and Pneumococcal Superinfection Induced by the Peptidoglycan of Lactobacillus rhamnosus CRL1505
    Authors: P Clua, M Tomokiyo, F Raya Tonet, MA Islam, V García Cas, G Marcial, S Salva, S Alvarez, H Takahashi, S Kurata, H Kitazawa, J Villena
    Cells, 2020;9(7):.
    Species: Mouse
    Sample Types: BALF
  5. IL-27 Is Essential for Suppression of Experimental Allergic Asthma by the TLR7/8 Agonist R848 (Resiquimod)
    Authors: Gesine Hansen
    J. Immunol., 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  6. Full Spectrum of LPS Activation in Alveolar Macrophages of Healthy Volunteers by Whole Transcriptomic Profiling
    PLoS ONE, 2016;11(7):e0159329.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. Andrographolide Restores Steroid Sensitivity To Block Lipopolysaccharide/IFN-?-Induced IL-27 and Airway Hyperresponsiveness in Mice
    J Immunol, 2016;196(11):4706-12.
    Species: Mouse
    Sample Types: BALF
  8. Interleukin-27 Mediates Susceptibility to Visceral Leishmaniasis by Suppressing the IL-17-Neutrophil Response
    Authors: Gustavo F S Quirino
    Infect Immun, 2016;0(0):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  9. TIR-domain-containing adapter-inducing interferon-beta (TRIF) regulates Th17-mediated intestinal immunopathology in colitis.
    Authors: Kanagavelu S, Flores C, Termini J, Riveron R, Romero L, Chung K, Ruiz J, Hyun J, Yuan X, Dagvadorj J, Fukata M
    Mucosal Immunol, 2015;8(2):296-306.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. IL-27 mediates the response to IFN-beta therapy in multiple sclerosis patients by inhibiting Th17 cells.
    Authors: Sweeney CM, Lonergan R, Basdeo SA, Kinsella K, Dungan LS, Higgins SC, Kelly PJ, Costelloe L, Tubridy N, Mills KH, Fletcher JM
    Brain Behav. Immun., 2010;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates

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Reviews for Mouse IL-27 p28/IL-30 DuoSet ELISA

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Mouse IL-27 p28/IL-30 DuoSet ELISA
By Anonymous on 08/10/2019
Sample Tested: Cell Lysates

Mouse IL-27 p28/IL-30 DuoSet ELISA
By Anonymous on 10/08/2018
Sample Tested: Serum