Human IL-4 QuantiGlo ELISA Kit

Name: Human IL-4 QuantiGlo ELISA Kit
Brand: R&D Systems
SKU: Q4000

Catalog #: Q4000 Datasheet / COA / SDS

Discontinued Product

Q4000 has been discontinued.

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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human IL-4 QuantiGlo ELISA Kit Summary

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Assay Length

5.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (150 uL), Serum (150 uL), EDTA Plasma (150 uL), Heparin Plasma (150 uL), Citrate Plasma (150 uL)

Sensitivity

1 pg/mL

Assay Range

1.6 - 5,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)

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.

Product Summary

The QuantiGlo Human IL-4 Chemiluminescent Immunoassay is a 5.5 hour solid phase ELISA designed to measure human 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 accurately quantitate recombinant human IL-4. Results obtained using natural human IL-4 showed dose response curves that were parallel to the standard curves obtained using the recombinant QuantiGlo 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) Four samples of known concentration were tested on one plate to assess intra-assay precision

Inter-Assay Precision (Precision between assays) Four 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	4	1	2	3	4
n	20	20	20	20	40	40	40	40
Mean (pg/mL)	2.38	10.1	125	1306	2.8	11.9	137	1400
Standard Deviation	0.21	0.85	8.13	100	0.41	1	7.2	60
CV%	8.8	8.4	6.5	7.7	14.6	8.4	5.3	4.3

Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma

	Intra-Assay Precision				Inter-Assay Precision
Sample	1	2	3	4	1	2	3	4
n	20	20	20	20	40	40	40	40
Mean (pg/mL)	3.63	18.9	177	1666	3.7	15.9	166	1547
Standard Deviation	0.34	0.56	8.39	78.1	0.43	1.3	13	117
CV%	9.4	3	4.7	4.7	11.6	8.2	7.8	7.6

Recovery

The recovery of human IL-4 spiked to three different levels throughout the range of the assay was evaluated.

Sample Type	Average % Recovery	Range %
Cell Culture Media (n=9)	103	87-115
Citrate Plasma (n=6)	91	83-100
EDTA Plasma (n=6)	92	78-106
Heparin Plasma (n=6)	92	82-102
Serum (n=6)	93	76-109

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of IL-4 in various matrices were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.\line

Scientific Data

N/A

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Product Datasheets

Product Datasheet

COA

SDS

Preparation and Storage

Storage

Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 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; IL4E12; 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.

Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

100 µL Assay Diluent

Add 100 µL of Assay Diluent to each well.

150 µL Standard, Control, or Sample

Add 150 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

200 µL Conjugate

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

200 µL Substrate Solution

Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20-40 minutes on the benchtop.
Determine the RLU of each well using a luminometer.