Human Interleukin 2 Receptor, IL2RA ELISA Kit

Sample

cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Species Reactivity

Human

Detection Method

sELISA

Intended Use

Quantitate Human IL2RA in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Contents of Kit

All reagents
Bring all reagents to room temperature (18-25°C) prior to use. Please DO NOT equilibrate unused plate well strips to room temperature. They should be sealed and stored in the original packaging. The assay can also be done at room temperature however we recommend doing it at 37°C for best consistency with our QC results. Also, the TMB incubation time estimate (15-25 min) is based on incubation at 37°C.
Wash buffer
Prepare 500 ml of Working Wash Buffer by diluting the supplied 20 ml of Wash Buffer (25 ×) with 480 ml of deionized or distilled water. If crystals have formed in the concentrate, warm to room temperature and mix it gently until crystals have completely dissolved.
Biotinylated Anti-Human IL2RA antibody
It is recommended to prepare this reagent immediately prior to use by diluting the Human IL2RA Biotinylated antibody (100×) 1:100 with Antibody Diluent. Prepare 100 μl by adding 1 μl of Biotinylated antibody (100×) to 99 μl of Antibody Diluent for each well. Mix gently and thoroughly and use within 2 hours of generation.
Avidin-Biotin-Peroxidase Complex
It is recommended to prepare this reagent immediately prior to use by diluting the Avidin-Biotin-Peroxidase Complex (100×) 1:100 with Avidin-Biotin-Peroxidase Diluent. Prepare 100 μl by adding 1 μl of Avidin-BiotinPeroxidase Complex (100x) to 99 μl of Avidin-Biotin-Peroxidase Diluent for each well. Mix gently and thoroughly and use within 2 hours of generation.
Human IL2RA Standard
It is recommended that the standards be prepared no more than 2 hours prior to performing the experiment. Use one 10 ng of lyophilized Human IL2RA standard for each experiment. Gently spin the vial prior to use. Reconstitute the standard to a stock concentration of 10 ng/ml using 1ml of sample diluent. Allow the standard to sit for a minimum of 10 minutes with gentle agitation prior to making dilutions.
Microplate
The included microplate is coated with capture antibodies and is ready-to-use. It does not require additional washing or blocking. The unused well strips should be sealed and stored in the original packaging.
Samples
Dilute the sample so that the expected range of concentrations fall within the detection range of this kit.
If the expected range of concentration is unknown, a pilot test should be conducted to decide the optimal dilution ratio for your samples.

Dilution of Human IL2RA Standard
1. Number tubes 1-8. Final Concentrations to be Tube # 1 –5,000.00 pg/ml, # 2 –2,500.00 pg/ml, # 3 – 1,250.00 pg/ml, # 4 – 625.00 pg/ml, # 5 –312.50 pg/ml, # 6 – 156.25 pg/ml, # 7 – 78.13 pg/ml, # 8 – Sample Diluent serves as the zero standard (0 pg/ml).
2. To generate standard #1, add 500 μl of the reconstituted standard stock solution of 10 ng/ml and 500 μl of sample diluent to tube #1 for a final volume of 1000 μl. Mix thoroughly.
3. Add 300 μl of sample diluent to tubes # 2-7.
4. To generate standard # 2, add 300 μl of standard # 1 from tube # 1 to tube # 2 for a final volume of 600 μl. Mix thoroughly.
5. To generate standard # 3, add 300 μl of standard # 2 from tube # 2 to tube # 3 for a final volume of 600 μl. Mix thoroughly.
6. Continue the serial dilution for tube # 4-7.

Storage

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Performance Characteristics

Technical Details
Capture/Detection Antibodies: The capture antibody is monoclonal antibody from mouse and the detection antibody is polyclonal antibody from goat.
Specificity: Natural and recombinant Human IL2RA.
Immunogen: Expression system for standard: NS0; Immunogen sequence: E22-C213.
Cross Reactivity: This kit is for the detection of Human IL2RA. No significant cross-reactivity or interference between IL2RA and its analogs was observed. This claim is limited by existing techniques therefore crossreactivity may exist with untested analogs.

Precision

We spend great efforts in documenting lot-to-lot variability and ensuring our assay kits produce robust data that are reproducible.
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 across assays): Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Detection Range

78 pg/ml - 5,000 pg/ml

Sensitivity

<5 pg/ml

Standard Curve

The highest O.D. value might be higher or lower than in the example. The experiment result is statistically significant if the highest O.D. value is no less than 1.0.

A standard curve is provided for demonstration only. A standard curve should be generated for each set of samples assayed.