Human IL-4 Quantikine HS ELISA Kit

R&D Systems | Catalog # HS400

R&D Systems
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Key Product Details

Assay Length

6.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (200 µL), Serum (200 µL)

Sensitivity

0.22 pg/mL

Assay Range

0.25-16 pg/mL (Cell Culture Supernates, Serum)
View all IL-4 ELISA Kits »

Product Summary for Human IL-4 Quantikine HS ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450 or 490nm (TMB or AP Substrate)

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

Interference observed with 1 or more available related molecules.

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 (pg/mL) 0.68 3.79 12.3 0.61 3.56 11.1
Standard Deviation 0.05 0.30 0.94 0.06 0.29 0.90
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 (pg/mL) 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 5.0 2.7 10.2 5.7 7.4

Recovery for Human IL-4 Quantikine HS ELISA Kit

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.

Human IL-4 High Sensitivity ELISA Linearity

Scientific Data Images for Human IL-4 Quantikine HS ELISA Kit

Human IL-4 High Sensitivity ELISA Cell Culture Supernate Standard Curve

Human IL-4 High Sensitivity ELISA Cell Culture Supernate Standard Curve

Human IL-4 High Sensitivity ELISA Serum Standard Curve

Human IL-4 High Sensitivity ELISA Serum 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 HS 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 HS ELISA Kit

For research use only

Citations for Human IL-4 Quantikine HS ELISA Kit

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Protocols

View specific protocols for Human IL-4 Quantikine HS ELISA Kit (HS400):

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
  3.   Add 50 µL of Assay Diluent to each well.
    4. 200 µL Standard, Control, or Sample
  4.   Add 200 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 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 4 times.
    8. 50 µL Substrate Solution
  8.   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.
    9. 50 µL Amplifier Solution
  9.   Add 50 µL Amplifier Solution to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
    10. 50 µL Stop Solution
  10. 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.

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

FAQs for Human IL-4 Quantikine HS 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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