Human IL-18 BPa DuoSet ELISA

Catalog # Availability Size / Price Qty
DY119
Ancillary Products Available
Human IL-18 BPa ELISA Standard Curve
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Product Details
Procedure
Citations (16)
FAQs
Supplemental Products
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Human IL-18 BPa DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
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 human IL-18 BPa. 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 ELISA.

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


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 equivalent

Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Blocking Buffer: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

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), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

 

Data Example

Human IL-18 BPa ELISA Standard Curve

Product Datasheets

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-18 BPa

Interleukin 18 binding proteins are secreted glycoproteins which antagonize IL-18 activity by binding IL-18 and blocking its biological activity. Alternative splicing of the IL-18 BP gene produces at least four isoforms that differ in affinity for IL-18. The IL-18 BP isoforms a and c each contain one immunoglobulin (Ig)-like C2-type domain while isoforms b and d lack a complete Ig-like domain.

Long Name:
Interleukin 18 Binding Protein Isoform a
Entrez Gene IDs:
10068 (Human); 84388 (Rat); 107126419 (Cynomolgus Monkey)
Alternate Names:
IL18 BPa; IL-18 BPa; IL18BP; IL-18BP; IL18BPa; interleukin 18 binding protein; interleukin-18-binding protein; MC51L-53L-54L homolog gene product; tadekinig-alfa

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody (to the working concentration stated in the product datasheet ) 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 each well of the microplate as recommended in the product datasheet. Incubate at room temperature for a minimum of 1 hour.

    Note: The recommended Reagent Diluent typically contains 1% BSA. Some DuoSet Development Kits require alternative blocking agents, or for plates to be blocked overnight with a higher percentage of BSA, please see the product datasheet for details.
     
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

 

PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.

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 (as recommended in the product datasheet), 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 Human IL-18 BPa 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.

16 Citations: Showing 1 - 10
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  1. Systemic inflammation is associated with malaria and preterm birth in women living with HIV on antiretrovirals and co-trimoxazole
    Authors: CR McDonald, AM Weckman, AL Conroy, P Olwoch, P Natureeba, MR Kamya, DV Havlir, G Dorsey, KC Kain
    Sci Rep, 2019;9(1):6758.
    Species: Human
    Sample Types: Plasma
  2. Resistance to visceral obesity is associated with increased locomotion in mice expressing an endothelial cell-specific fibroblast growth factor 1 transgene
    Authors: T Keeley, A Kirov, WY Koh, V Demambro, I Bergquist, J Cotter, P Caradonna, ME Siviski, B Best, T Henderson, CJ Rosen, L Liaw, I Prudovsky, DJ Small
    Physiol Rep, 2019;7(7):e14034.
    Species: Human
    Sample Types: Serum
  3. Early malaria infection, dysregulation of angiogenesis, metabolism and inflammation across pregnancy, and risk of preterm birth in Malawi: A cohort study
    Authors: RE Elphinston, AM Weckman, CR McDonald, V Tran, K Zhong, M Madanitsa, L Kalilani-P, C Khairallah, SM Taylor, SR Meshnick, V Mwapasa, FO Ter Kuile, AL Conroy, KC Kain
    PLoS Med., 2019;16(10):e1002914.
    Species: Human
    Sample Types: Plasma
  4. Increased interleukin 18 activity in adolescents with early-onset psychosis is associated with cortisol and depressive symptoms
    Authors: K Wedervang-, S Friis, V Lonning, RE Smelror, C Johannesse, EJ Reponen, SH Lyngstad, T Lekva, P Aukrust, T Ueland, OA Andreassen, I Agartz, AM Myhre
    Psychoneuroendocrinology, 2019;0(0):104513.
    Species: Human
    Sample Types: Serum
  5. Adipose PTEN regulates adult adipose tissue homeostasis and redistribution via a PTEN-leptin-sympathetic loop
    Authors: W Huang, NJ Queen, TB McMurphy, S Ali, L Cao
    Mol Metab, 2019;30(0):48-60.
    Species: Mouse
    Sample Types: Serum
  6. Asymmetric dimethylarginine serum levels are associated with early mortality after allogeneic stem cell transplantation
    Authors: A Radujkovic, H Dai, L Kordelas, D Beelen, SP Rachakonda, C Müller-Tid, R Kumar, P Dreger, T Luft
    Haematologica, 2018;0(0):.
    Species: Human
    Sample Types: Serum
  7. Production and regulation of interleukin-1 family cytokines at the materno-fetal interface
    Authors: LM Scott, AH Bryant, A Rees, B Down, RH Jones, CA Thornton
    Cytokine, 2017;0(0):.
    Species: Human
    Sample Types: Tissue Culture Supernates
  8. Elevated hepatic DPP4 activity promotes insulin resistance and non-alcoholic fatty liver disease
    Authors: C Baumeier, L Schlüter, S Saussentha, T Laeger, M Rödiger, SA Alaze, L Fritsche, HU Häring, N Stefan, A Fritsche, RW Schwenk, A Schürmann
    Mol Metab, 2017;6(10):1254-1263.
    Species: Mouse
    Sample Types: Plasma
  9. Inflammatory and Angiogenic Factors at Mid-Pregnancy Are Associated with Spontaneous Preterm Birth in a Cohort of Tanzanian Women.
    Authors: McDonald C, Darling A, Conroy A, Tran V, Cabrera A, Liles W, Wang M, Aboud S, Urassa W, Fawzi W, Kain K
    PLoS ONE, 2015;10(8):e0134619.
    Species: Human
    Sample Types: Plasma
  10. Human papillomavirus E7 oncoprotein increases production of the anti-inflammatory interleukin-18 binding protein in keratinocytes.
    Authors: Richards, Kathryn, Doble, Rosella, Wasson, Christop, Haider, Mohammed, Blair, G Eric, Wittmann, Miriam, Macdonald, Andrew
    J Virol, 2014;88(8):4173-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Host biomarkers distinguish dengue from leptospirosis in Colombia: a case-control study.
    Authors: Conroy A, Gelvez M, Hawkes M, Rajwans N, Liles W, Villar-Centeno L, Kain K
    BMC Infect Dis, 2014;14(0):35.
    Species: Human
    Sample Types: Serum
  12. Quiescent innate response to infective filariae by human Langerhans cells suggests a strategy of immune evasion.
    Authors: Boyd A, Bennuru S, Wang Y, Sanprasert V, Law M, Chaussabel D, Nutman T, Semnani R
    Infect Immun, 2013;81(5):1420-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Impact of vitamin D supplementation on markers of inflammation in adults with cystic fibrosis hospitalized for a pulmonary exacerbation.
    Eur J Clin Nutr, 2012;66(9):1072-4.
    Species: Human
    Sample Types: Serum
  14. IL-18 mediates proapoptotic signaling in renal tubular cells through a Fas ligand-dependent mechanism.
    Authors: Zhang H, Hile KL, Asanuma H
    Am. J. Physiol. Renal Physiol., 2011;301(1):F171-8.
    Species: Human
    Sample Types: Serum
  15. IL-8 dictates glycosaminoglycan binding and stability of IL-18 in cystic fibrosis.
    Authors: Reeves EP, Williamson M, Byrne B, Bergin DA, Smith SG, Greally P, O'Kennedy R, O'Neill SJ, McElvaney NG
    J. Immunol., 2010;184(3):1642-52.
    Species: Human
    Sample Types: Serum
  16. Defective phosphorylation of interleukin-18 receptor beta causes impaired natural killer cell function in systemic-onset juvenile idiopathic arthritis.
    Authors: De Jager W, Vastert SJ, Beekman JM, Wulffraat NM, Kuis W, Coffer PJ, Prakken BJ
    Arthritis Rheum., 2009;60(9):2782-93.
    Species: Human
    Sample Types: Plasma

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Human IL-18 BPa DuoSet ELISA
By Anonymous on 09/03/2018
Application: Sample Tested: EDTA Plasma

We ran EDTA-samples at a 1:20 dilution for this assay with an intra-variation of 2.2%