Human IL-5 Quantikine ELISA Kit

R&D Systems | Catalog # D5000B

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL), Urine (100 µL)

Sensitivity

1.08 pg/mL

Assay Range

3.9-250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
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Product Summary for Human IL-5 Quantikine ELISA Kit

The Quantikine Human IL-5 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-5 levels in cell culture supernates, serum, plasma, and urine. It contains Sf 21-expressed recombinant human IL-5 and antibodies raised against the recombinant factor and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-5 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural IL-5.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human IL-5

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.

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, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 28.1 89.3 170 31.2 92.6 174
Standard Deviation 0.89 2.0 7.27 2.5 4.2 8.7
CV% 3.2 2.2 4.2 8.0 4.5 5.0

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) 31.3 95.6 173 32.9 97.9 180
Standard Deviation 1.4 3.5 6.5 4.0 6.7 8.4
CV% 4.5 3.7 3.8 12.2 6.8 4.7

Recovery for Human IL-5 Quantikine ELISA Kit

The recovery of human IL-5 spiked to different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 101 95-107
EDTA Plasma (n=4) 104 86-118
Heparin Plasma (n=4) 101 87-112
Serum (n=4) 104 88-116
Urine (n=4) 96 91-106

Linearity

To assess the linearity of the assay, five samples were spiked with high concentrations of IL-5 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human IL-5 ELISA Linearity

Scientific Data Images for Human IL-5 Quantikine ELISA Kit

Human IL-5 ELISA Cell Culture Supernate/Urine Standard Curve

Human IL-5 ELISA Cell Culture Supernate/Urine Standard Curve

Human IL-5 ELISA Serum/Plasma Standard Curve

Human IL-5 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-5

IL-5 (Interleukin-5) is a secreted cytokine that is expressed as a covalent dimer. It is produced by activated Th2 cells, eosinophils, mast cells, and iNKT cells and promotes eosinophilic and basophilic differentiation and activation. IL-5 signals through a receptor complex containing IL-5 R alpha and the Common beta Chain/CD131 on eosinophils and basophils. Naturally occurring soluble forms of IL-5 R alpha retain the ability to bind IL-5 and limit its bioavailability.

Long Name

Interleukin 5

Alternate Names

BCDF mu, BCGFII, EDF, Eo-CSF, IL5, TRF

Entrez Gene IDs

3567 (Human); 16191 (Mouse); 24497 (Rat); 397409 (Porcine); 280825 (Bovine); 403790 (Canine); 100034199 (Equine); 493803 (Feline)

Gene Symbol

IL5

Additional IL-5 Products

Product Documents for Human IL-5 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-5 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-5 Quantikine ELISA Kit

Customer Reviews for Human IL-5 Quantikine ELISA Kit (3)

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  • Human IL-5 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 10/29/2020
    Easy to use, Good results
    Human IL-5 Quantikine ELISA Kit D5000B
  • Human IL-5 Quantikine ELISA Kit
    Name: Evan Williams
    Sample Tested: EDTA Plasma
    Verified Customer | Posted 10/28/2020
    Kit worked well, however human plasma had undetectable levels of IL-5
    Human IL-5 Quantikine ELISA Kit D5000B
  • Human IL-5 Quantikine ELISA Kit
    Name: Alisha Freeman
    Sample Tested: Calu-6 human lung carcinoma cell line
    Verified Customer | Posted 10/10/2017

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Protocols

View specific protocols for Human IL-5 Quantikine ELISA Kit (D5000B):

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. 100 µL Standard, Control, or Sample
  4.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  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 on the shaker.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. 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.

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FAQs

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