Human IL-4 Quantikine HS ELISA Kit

  (21 citations)     
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Product Details
Assay Procedure
Citations (21)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    6.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (200 uL), Serum (200 uL)
  • Sensitivity
    0.22 pg/mL
  • Assay Range
    0.3 - 16 pg/mL (Cell Culture Supernates, Serum)
  • Specificity
    Natural and recombinant human IL-4
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Control Available
QC41 , Quantikine Immunoassay Control Group 10 - Please Inquire
Product Summary
The Quantikine HS Human IL-4 Immunoassay is a 6.5 hour solid phase ELISA designed to measure IL-4 levels in cell culture supernates and serum. It contains E. coli-expressed recombinant human IL-4 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant human IL-4 accurately. Results obtained using natural human IL-4 showed linear curves that were parallel to the standard curves obtained using the Quantikine HS kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-4.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 0.68 3.79 12.3 0.61 3.56 11.1
Standard Deviation 0.05 0.3 0.94 0.06 0.29 0.9
CV% 7.4 7.9 7.6 9.8 8.1 8.1

Serum
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 1.75 5.56 10.7 0.59 3.32 10.7
Standard Deviation 0.05 0.28 0.29 0.06 0.19 0.79
CV% 2.9 5 2.7 10.2 5.7 7.4

Recovery

The recovery of IL-4 was determined by spiking to levels throughout the range of the assay in various matrices.

Sample Type Average % Recovery Range %
Cell Culture Media (n=9) 106 87-118
Serum (n=4) 102 95-108
Linearity
To assess linearity of the assay, the following samples, spiked with high concentrations of IL-4, were diluted with the appropriate Calibrator Diluent and then assayed.
 IL-4 [HRP]
Product Datasheets

Certificate of Analysis Lookup

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Note: Certificate of Analysis not available for kit components.

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: IL-4
IL-4 (Interleukin-4) is a cytokine that is primarily expressed by Th2-biased CD4+ T cells, mast cells, basophils, and eosinophils. It promotes cell proliferation, survival, and immunoglobulin class switch to IgG4 and IgE in human B cells, acquisition of the Th2 phenotype by naïve CD4+ T cells, priming and chemotaxis of mast cells, eosinophils, and basophils, and the proliferation and activation of epithelial cells. IL-4 plays a dominant role in the development of allergic inflammation and asthma. The type I receptor for IL-4, which is expressed on hematopoietic cells, is a heterodimer of the ligand binding IL-4 R? and the common ? chain (a shared subunit of the receptors for IL-2, -7, -9, -15, and -21). The type II receptor on nonhematopoietic cells consists of IL-4 R? and IL-13 R?1. The type II receptor also transduces IL-13 mediated signals.
  • Long Name:
    Interleukin 4
  • Entrez Gene IDs:
    3565 (Human); 16189 (Mouse); 287287 (Rat); 397225 (Porcine); 280824 (Bovine); 403785 (Canine); 574281 (Primate); 100302454 (Rabbit)
  • Alternate Names:
    B cell growth factor 1; BCDF; B-cell stimulatory factor 1; BCGF1; BCGF-1; binetrakin; BSF1; BSF-1; IL4; IL-4; IL-4B_cell stimulatory factor 1; interleukin 4; interleukin-4; Lymphocyte stimulatory factor 1; MGC79402; pitrakinra
Related Research Areas
Assay Procedure
Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 200 µL Standard, Control, or Sample
  6.   Add 200 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 50 µL Substrate Solution
  12.   Add 50 µL Substrate Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour. Do not wash the plate.

  13. 50 µL Amplifier Solution
  14.   Add 50 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.

  15. 50 µL Stop Solution
  16. Add 50 µL of Stop Solution to each well. Read at 490 nm within 30 minutes. Set wavelength correction to 650 nm or 690 nm.
Citations:

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.

21 Citations: Showing 1 - 10
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Species
Sample Type
  1. Effects of interval and continuous exercise training on CD4 lymphocyte apoptotic and autophagic responses to hypoxic stress in sedentary men.
    Authors: Weng T, Huang S, Chuang Y, Wang J
    PLoS ONE, 2013;8(11):e80248.
    Species: Human
    Sample Type: Plasma
  2. Leptin enhances survival and induces migration, degranulation, and cytokine synthesis of human basophils.
    Authors: Suzukawa M, Nagase H, Ogahara I, Han K, Tashimo H, Shibui A, Koketsu R, Nakae S, Yamaguchi M, Ohta K
    J. Immunol., 2011;186(9):5254-60.
    Species: Human
    Sample Type: Cell Culture Supernates
  3. Subnormal cytokine profile in the tear fluid of keratoconus patients.
    Authors: Jun AS, Cope L, Speck C, Feng X, Lee S, Meng H, Hamad A, Chakravarti S
    PLoS ONE, 2011;6(1):e16437.
    Species: Human
    Sample Type: Tears
  4. Sensitive plasma protein analysis by microparticle-based proximity ligation assays.
    Authors: Darmanis S, Nong RY, Hammond M, Gu J, Alderborn A, Vanelid J, Siegbahn A, Gustafsdottir S, Ericsson O, Landegren U, Kamali-Moghaddam M
    Mol. Cell Proteomics, 2010;9(2):327-35.
    Species: Human
    Sample Type: Plasma
  5. Urinary proinflammatory cytokine response in renal transplant recipients with polyomavirus BK viruria.
    Authors: Sadeghi M, Daniel V, Schnitzler P, Lahdou I, Naujokat C, Zeier M, Opelz G
    Transplantation, 2009;88(9):1109-16.
    Species: Human
    Sample Type: Urine
  6. Detection of pancreatic cancer using antibody microarray-based serum protein profiling.
    Authors: Ingvarsson J, Wingren C, Carlsson A, Ellmark P, Wahren B, Engstrom G, Harmenberg U, Krogh M, Peterson C, Borrebaeck CA
    Proteomics, 2008;8(11):2211-9.
    Species: Human
    Sample Type: Serum
  7. Anti-inflammatory cytokines in gingival crevicular fluid in patients with periodontitis and rheumatoid arthritis: a preliminary report.
    Authors: Bozkurt FY, Yetkin Ay Z, Berker E, Tepe E, Akkus S
    Cytokine, 2006;35(3):180-5.
    Species: Human
    Sample Type: Gingival Crevicular Fluid (GCF)
  8. Expression of interleukin-18 and caspase-1 in cutaneous T-cell lymphoma.
    Authors: Yamanaka K, Clark R, Dowgiert R, Hurwitz D, Shibata M, Rich BE, Hirahara K, Jones DA, Eapen S, Mizutani H, Kupper TS
    Clin. Cancer Res., 2006;12(2):376-82.
    Species: Human
    Sample Type: Plasma
  9. Dysregulated monocyte iron homeostasis and erythropoietin formation in patients with anemia of chronic disease.
    Authors: Theurl I, Mattle V, Seifert M, Mariani M, Marth C, Weiss G
    Blood, 2006;107(10):4142-8.
    Species: Human
    Sample Type: Serum
  10. Th2 predominance and CD8+ memory T cell depletion in patients with severe acute respiratory syndrome.
    Authors: Huang JL, Huang J, Duan ZH, Wei J, Min J, Luo XH, Li JG, Tan WP, Wu LZ, Liu RY, Li Y, Shao J, Huang BJ, Zeng YX, Huang W
    Microbes Infect., 2005;7(3):427-36.
    Species: Human
    Sample Type: Serum
  11. Cytokines and adhesion molecules in multiple sclerosis patients treated with interferon-beta1b.
    Authors: Jensen J, Krakauer M, Sellebjerg F
    Cytokine, 2005;29(1):24-30.
    Species: Human
    Sample Type: Plasma
  12. Correlations between IL-4, IL-12 levels and CCL2, CCL5 levels in serum and cerebrospinal fluid of multiple sclerosis patients.
    Authors: Bartosik-Psujek H, Stelmasiak Z
    J Neural Transm, 2005;112(6):797-803.
    Species: Human
    Sample Type: Serum
  13. Strong inflammatory cytokine response in male and strong anti-inflammatory response in female kidney transplant recipients with urinary tract infection.
    Authors: Sadeghi M, Daniel V, Naujokat C, Wiesel M, Hergesell O, Opelz G
    Transpl. Int., 2005;18(2):177-85.
    Species: Human
    Sample Type: Plasma
  14. Antifungal immune reactivity in nasal polyposis.
    Authors: Pitzurra L, Bellocchio S, Nocentini A, Bonifazi P, Scardazza R, Gallucci L, Stracci F, Simoncelli C, Bistoni F, Romani L
    Infect. Immun., 2004;72(12):7275-81.
    Species: Human
    Sample Type: Nasal Lavage
  15. Exhaled breath condensate in children: pearls and pitfalls.
    Authors: Rosias PP, Dompeling E, Hendriks HJ, Heijnens JW, Donckerwolcke RA, Jobsis Q
    Pediatr Allergy Immunol, 2004;15(1):4-19.
    Species: Human
    Sample Type: Exhaled Breath Condensate (EBC)
  16. Immunodetection of a hepatitis C virus (HCV) antigen and Thl/Th2 cytokines in cerebrospinal fluid of meningitis patients.
    Authors: Attallah AM, Ibrahim GG
    J Immunoassay Immunochem, 2004;25(4):313-20.
    Species: Human
    Sample Type: Serum
  17. A double-blind, placebo-controlled immunotherapy dose-response study with standardized cat extract.
    Authors: Ewbank PA, Murray J, Sanders K, Curran-Everett D, Dreskin S, Nelson HS
    J. Allergy Clin. Immunol., 2003;111(1):155-61.
    Species: Human
    Sample Type: Nasal Fluid
  18. Cytokine profile in Behcet's disease patients. Relationship with disease activity.
    Authors: Hamzaoui K, Hamzaoui A, Guemira F, Bessioud M, Hamza M, Ayed K
    Scand. J. Rheumatol., 2002;31(4):205-10.
    Species: Human
    Sample Type: Serum
  19. Modulation of human immune response by Echinococcus granulosus antigen B and its possible role in evading host defenses.
    Authors: Rigano R, Profumo E, Bruschi F, Carulli G, Azzara A, Ioppolo S, Buttari B, Ortona E, Margutti P, Teggi A, Siracusano A
    Infect. Immun., 2001;69(1):288-96.
    Species: Human
    Sample Type: Cell Culture Supernates
  20. Low levels of interferon-gamma in nasal fluid accompany raised levels of T-helper 2 cytokines in children with ongoing allergic rhinitis.
    Authors: Benson M, Strannegard IL, Wennergren G, Strannegard O
    Pediatr Allergy Immunol, 2000;11(1):20-8.
    Species: Human
    Sample Type: Nasal Lavage
  21. Cytokine profiles in parotid saliva from HIV-1-infected individuals: changes associated with opportunistic infections in the oral cavity.
    Authors: Black, K P, Merrill, K W, Jackson, S, Katz, J
    Oral Microbiol Immunol, 2000;15(2):74-81.
    Species: Human
    Sample Type: Saliva
Expand to show all Citations

FAQs

  1. Why is plasma a validated sample type for the Human IL-4 Quantikine® ELISA (Catalog # D4050), but not in the high sensitivity version (Catalog # HS400)?

    • The recovery and linearity obtained with plasma samples assayed with the high sensitivity kit HS400 show too much variability and did not meet our specifications, unlike the D4050 which shows reliable recovery and linearity data with plasma samples.

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