Human CD25/IL-2R alpha Quantikine ELISA Kit

R&D Systems | Catalog # DR2A00

Quantikine Quickit version available - results in 90 minutes (QK0223).
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Key Product Details

Assay Length

3.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (10 µL), Serum (13 µL), EDTA Plasma (13 µL), Heparin Plasma (13 µL)

Sensitivity

10 pg/mL

Assay Range

78-5000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Human CD25/IL-2R alpha Quantikine ELISA Kit

The Quantikine Human IL-2 sR alpha Immunoassay is a 3.5 hour solid phase ELISA designed to measure IL-2 sR alpha in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human IL-2 sR alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IL-2 sR alpha showed linear 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-2 sR alpha.

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-2 sR alpha

Cross-reactivity

Cross-reactivity observed with 1 or more available related molecules. Cross-species reactivity not tested.

Interference

No significant interference observed with available related molecules.

Sample Values

Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence of human IL-2 R alpha in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL)
Serum (n=55)1055458-1997365
EDTA plasma (n=55)1062410-2623424
Heparin plasma (n=55)1020430-2082365

Cell Culture Supernates - Human peripheral blood mononuclear cells (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cell were cultured unstimulated or stimulated with 10 μg/mL of PHA for 1, 3, and 5 days. Aliquots of the cell culture supernates were removed and assayed for levels of human IL-2 Ra.

ConditionDay 1Day 3Day 5
Unstimulated245459645
Stimulated 383110,0017765



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, 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) 207 613 2357 188 630 2615
Standard Deviation 12.7 37.5 108 13.6 38.1 183
CV% 6.1 6.1 4.6 7.2 6.0 7.0

Recovery for Human CD25/IL-2R alpha Quantikine ELISA Kit

The recovery of IL-2 sR alpha spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 105 93-118
EDTA Plasma (n=5) 103 91-113
Heparin Plasma (n=5) 104 98-113
Serum (n=5) 105 99-116

Linearity

To assess the linearity of the assay, five samples containing or spiked with high concentrations of IL-2 sR alpha in various matrices were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human IL2RA ELISA Linearity

Scientific Data Images for Human CD25/IL-2R alpha Quantikine ELISA Kit

Human IL2RA ELISA Standard Curve

Human IL2RA ELISA 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: CD25/IL-2R alpha

CD25/IL-2 receptor alpha is a transmembrane protein that associates with IL-2R beta/CD122 and the Common gamma Chain/CD132 on activated T cells and regulatory T cells (Treg). The Common gamma chain is a shared subunit in the receptors for IL-4, -7, -9, -15, and -21. IL-2R alpha is required for mediating IL-2 induced effects including T cell proliferation, activation induced cell death (AICD) of autoreactive naive T cells, and the development of CD4+CD25+ Treg which contribute to peripheral T cell homeostasis. A soluble form of IL-2R alpha can be generated by proteolytic cleavage of the cell surface receptor.

Long Name

Interleukin 2 Receptor alpha

Alternate Names

CD25, IL-2 R alpha, IL-2Ra, IL2R alpha, IL2RA

Entrez Gene IDs

3559 (Human); 16184 (Mouse); 25704 (Rat); 403870 (Canine); 102123605 (Cynomolgus Monkey)

Gene Symbol

IL2RA

Additional CD25/IL-2R alpha Products

Product Documents for Human CD25/IL-2R alpha 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 CD25/IL-2R alpha 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 CD25/IL-2R alpha Quantikine ELISA Kit

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Protocols

View specific protocols for Human CD25/IL-2R alpha Quantikine ELISA Kit (DR2A00):

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. Proceed to the next step. Do not incubate or wash.
    5. 100 µL Conjugate
  5.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 3 hours.
  6.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    7. 200 µL Substrate Solution
  7.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    8. 50 µL Stop Solution
  8.   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 CD25/IL-2R alpha Quantikine ELISA Kit

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  • Q: Does the Human CD25/IL-2R alpha Quantikine ELISA Kit cross react with cyno samples?

    A: We have not tested animal samples in this kit. We are aware that customers have cited the use of this kit with cyno samples. See the Citations tab. Additional sample validation by the investigator may be required.

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