Human IL-4 Quantikine ELISA Kit

R&D Systems | Catalog # D4050

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Citrate Plasma (50 µL)

Sensitivity

10 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human IL-4 Quantikine ELISA Kit

The Quantikine Human IL-4 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-4 levels in cell culture supernates, serum, and plasma. 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 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 human IL-4.

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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

No significant interference observed with available related molecules.

精度

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 40 40 40
Mean (pg/mL) 159 538 1092 169 503 1032
Standard Deviation 11.4 27.8 67.2 16.8 49.3 83.0
CV% 7.2 5.2 6.2 9.9 9.8 8.0

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 170 510 1110 132 411 865
Standard Deviation 6.7 17.5 58.5 9.8 32.1 52.8
CV% 3.9 3.4 5.3 7.4 7.8 6.1

Recovery for Human IL-4 Quantikine ELISA Kit

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=8) 98 90-109
Citrate Plasma (n=10) 94 85-109
EDTA Plasma (n=10) 100 90-123
Heparin Plasma (n=10) 95 84-121
Serum (n=10) 98 82-111

Linearity

To assess the linearity of the assay, the following biological samples, spiked with high concentrations of IL-4, were diluted with the appropriate Calibrator Diluent and then assayed.

Human IL-4 ELISA Linearity

Scientific Data Images for Human IL-4 Quantikine ELISA Kit

Human IL-4 ELISA Cell Culture Supernate Standard Curve

Human IL-4 ELISA Cell Culture Supernate Standard Curve

Human IL-4 ELISA Serum/Plasma Standard Curve

Human IL-4 ELISA Serum/Plasma Standard Curve

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-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 alpha and the common gamma 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 alpha and IL-13 R alpha 1. The type II receptor also transduces IL-13 mediated signals.

Long Name

Interleukin 4

Alternate Names

BCDF, BCGF-1, BCGF1, BSF-1, BSF1, IL4, IL4E12

Entrez Gene IDs

3565 (Human); 16189 (Mouse); 287287 (Rat); 397225 (Porcine); 280824 (Bovine); 403785 (Canine); 751514 (Feline); 574281 (Primate); 100302454 (Rabbit)

Gene Symbol

IL4

Additional IL-4 Products

Product Documents for Human IL-4 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human IL-4 Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human IL-4 Quantikine ELISA Kit

Customer Reviews for Human IL-4 Quantikine ELISA Kit (3)

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  • Human IL-4 Quantikine ELISA Kit
    Name: Prasanna Ekambaram
    Sample Tested: T cells and Cell culture supernatant
    Verified Customer | Posted 09/05/2024
  • Human IL-4 Quantikine ELISA Kit
    Name: Alisha Freeman
    Sample Tested: 293T human embryonic kidney cell line
    Verified Customer | Posted 07/12/2018
  • Human IL-4 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: CHO Chinese hamster ovary cell line
    Verified Customer | Posted 04/24/2018

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Protocols

View specific protocols for Human IL-4 Quantikine ELISA Kit (D4050):

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. 100 µL Assay Diluent
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 3 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Human IL-4 Quantikine ELISA Kit

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    • Q: 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)?
      A: 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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