Rat IL-4 Quantikine ELISA Kit

  (10 citations)     
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Assay Procedure
Citations (10)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (50 uL)
  • Sensitivity
    5 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Serum)
  • Specificity
    Natural and recombinant rat IL-4
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Rat IL-4 Immunoassay is a 4.5 hour solid phase ELISA designed to measure rat IL-4 levels in cell culture supernates and serum. It contains E. coli-expressed recombinant rat IL-4 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant rat IL-4 accurately. Results obtained using natural rat IL-4 showed dose response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural rat 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, Serum
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 48.2 111 721 47.7 112 742
Standard Deviation 2.5 4.9 23.6 3.9 6.1 31.8
CV% 5.2 4.4 3.3 8.2 5.4 4.3

Recovery

The recovery of rat IL-4 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 110 104-117
Serum (n=7) 94 88-101
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of rat IL-4 in each matrix were diluted with Calibrator Diluent and then assayed.
 IL-4 [HRP]
Product Datasheets

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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. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 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.

10 Citations: Showing 1 - 10
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Species
Sample Type
  1. Age-, Gender-, andin VivoDifferent Doses of Isoproterenol Modifyin VitroAortic Vasoreactivity and Circulating VCAM-1
    Authors: B Nieto-Lima, A Cano-Martí, ME Rubio-Ruiz, I Pérez-Torr, V Guarner-La
    Front Physiol, 2018;9(0):20.
    Species: Mouse
    Sample Type: Plasma
  2. Topical use and systemic action of green and roasted coffee oils and ground oils in a cutaneous incision model in rats (Rattus norvegicus albinus)
    Authors: BG Lania, J Morari, AL Souza, MND Silva, AR de Almeida, G Veira-Dami, SM Alegre, CL César, LA Velloso, ML Cintra, NB Maia, PENF Velho
    PLoS ONE, 2017;12(12):e0188779.
    Species: Rat
    Sample Type: Serum
  3. Association of C-Type Lectin Mincle with Fc?RI?? Subunits Leads to Functional Activation of RBL-2H3 Cells through Syk
    Authors: C Honjoh, K Chihara, H Yoshiki, S Yamauchi, K Takeuchi, Y Kato, Y Hida, T Ishizuka, K Sada
    Sci Rep, 2017;7(0):46064.
    Species: Rat
    Sample Type: Cell Culture Supernates
  4. The effects of low-dose ionizing radiation in the activated rat basophilic leukemia (RBL-2H3) mast cells.
    Authors: Joo H, Nam S, Yang K, Kim C, Jin Y, Kim J
    J Biol Chem, 2012;287(33):27789-95.
    Species: Rat
    Sample Type: Cell Culture Supernates
  5. Comprehensive gene expression profiling of rat lung reveals distinct acute and chronic responses to cigarette smoke inhalation.
    Authors: Stevenson CS, Docx C, Webster R, Battram C, Hynx D, Giddings J, Cooper PR, Chakravarty P, Rahman I, Marwick JA, Kirkham PA, Charman C, Richardson DL, Nirmala NR, Whittaker P, Butler K
    Am. J. Physiol. Lung Cell Mol. Physiol., 2007;293(5):L1183-93.
    Species: Rat
    Sample Type: BALF
  6. Cellular recruitment and cytokine generation in a rat model of allergic lung inflammation are differentially modulated by progesterone and estradiol.
    Authors: de Oliveira AP, Domingos HV, Cavriani G, Damazo AS, Dos Santos Franco AL, Oliani SM, Oliveira-Filho RM, Vargaftig BB, de Lima WT
    Am. J. Physiol., Cell Physiol., 2007;293(3):C1120-8.
    Species: Rat
    Sample Type: BALF
  7. Suppression of autoimmune retinal disease by lovastatin does not require Th2 cytokine induction.
    Authors: Gegg ME, Harry R, Hankey D, Zambarakji H, Pryce G, Baker D, Adamson P, Calder V, Greenwood J
    J. Immunol., 2005;174(4):2327-35.
    Species: Rat
    Sample Type: Cell Culture Supernates
  8. Insulin attenuates the systemic inflammatory response in endotoxemic rats.
    Authors: Jeschke MG, Klein D, Bolder U, Einspanier R
    Endocrinology, 2004;145(9):4084-93.
    Species: Rat
    Sample Type: Tissue Homogenates
  9. Insulin treatment improves hepatic morphology and function through modulation of hepatic signals after severe trauma.
    Authors: Schubert T, Horch RE
    Ann. Surg., 2004;240(2):340-9.
    Species: Rat
    Sample Type: Tissue Homogenates
  10. Local anticandidal immune responses in a rat model of vaginal infection by and protection against Candida albicans.
    Authors: de Bernardis F, Santoni G, Boccanera M, Spreghini E, Adriani D, Morelli L, Cassone A
    Infect. Immun., 2000;68(6):3297-304.
    Species: Rat
    Sample Type: Vaginal Fluid

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