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Product Info Summary
| SKU: | EK0400 |
|---|---|
| Size: | 96 wells/kit, with removable strips. |
| Reactive Species: | Human |
| Application: | ELISA |
| Sample Types: | cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). |
Product info
Product Name
Human CD25/IL-2sR Alpha ELISA Kit PicoKine®
View all CD25/IL-2R alpha ELISA kits
SKU/Catalog Number
EK0400
Size
96 wells/kit, with removable strips.
*Question: How many samples can I assay/run in this kit?
Description
Human CD25/IL-2sR Alpha ELISA Kit PicoKine® (96 Tests). Quantitate Human IL2RA in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 5pg/ml. The brand Picokine indicates this is a premium quality ELISA kit. Each Picokine kit delivers precise quantification, high sensitivity, and excellent reproducibility. Only our most reliable and effective kits qualify as Picokine, guaranteeing top-tier results for your assays.
Storage & Handling
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.)
Cite This Product
Human CD25/IL-2sR Alpha ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0400)
Clonality of Antibodies
See Datasheet for details
Standard Protein
Expression system for standard: NS0; Immunogen sequence: E22-C213
Sensitivity
<5 pg/ml
Assay Range
78 pg/ml - 5,000 pg/ml
Standard Dilution Instructions
See datasheet of EK0400 for more details
Cross-reactivity
There is no detectable cross-reactivity with other relevant proteins.
Reactive Species
EK0400 is reactive to IL2RA in Human samples
Validated Sample Types
cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
Application Guarantee
EK0400 is guaranteed for ELISA in Human by Boster Guarantee
See how Boster Bio validate our ELISA kits: ELISA Validation Information
Background of CD25/IL-2R alpha
CD25 is the alpha chain of the IL-2 receptor. It is a type I transmembrane protein present on activated T cells, activated B cells, some thymocytes, myeloid precursors, and oligodendrocytes that associates with CD122 to form a heterodimer that can act as a high-affinity receptor for IL-2. The IL2R gene was mapped to 10p15-p14. CD25 is expressed in most B-cell neoplasms, some acute nonlymphocytic leukemias, neuroblastomas, and tumor infiltrating lymphocytes. Its soluble form, called sIL-2R may be elevated in these diseases and is occasionally used to track disease progression.
Kit Components
| Catalog Number | Description | Quantity |
|---|---|---|
| EK0400-CAP | Anti-Human IL2RA Pre-coated 96-well strip microplate | 1 |
| EK0400-ST | Human IL2RA Standard | 2 vials, 10 ng/tube |
| EK0400-DA | Human IL2RA Biotinylated antibody (100x) | 100ul |
| AR1103 | Avidin-Biotin-Peroxidase Complex (100x) | 100ul |
| AR1106-1 | Sample Diluent | 30ml |
| AR1106-2 | Antibody Diluent | 12ml |
| AR1106-3 | Avidin-Biotin-Peroxidase Diluent | 12ml |
| AR1104 | Color Developing Reagent (TMB) | 10ml |
| AR1105 | Stop Solution | 10ml |
| AR1106-5 | Wash Buffer (25x) | 20ml |
| PLA-SEA | Adhesive plate sealers | 4 |
*The kit components are not available for individual purchase.
Materials Required But Not Included In Kit
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
Data Examples, Quality Control Data & Sample Dilution
Validation Standard Curve O.D. At 450nm
| Concentration (pg/ml) | 0 | 78 | 156 | 313 | 625 | 1250 | 2500 | 5000 |
| O.D. | 0.016 | 0.251 | 0.435 | 0.770 | 1.108 | 1.479 | 1.853 | 2.049 |
Data Example Images
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Human CD25/IL-2sR alpha PicoKine ELISA Kit standard curve
Recommended Sample Dilution Ratios
According to our internal validation assays using this ELISA kit, to detect CD25/IL-2R alpha, Dilution ratio of 1:50, concentration in serum and plasma is 670-1178 pg/ml..
Intra Assay Consistency & Inter Assay Consistency
We measured random samples of Human CD25/IL-2sR Alpha ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 140 | 719 | 2353 | 141 | 785 | 2310 |
| Standard deviation | 6.58 | 41.7 | 94.12 | 7.61 | 47.1 | 97.02 |
| CV (%) | 4.7% | 5.8% | 4% | 5.4% | 6% | 4.2% |
Reproducibility
We ensure reproducibility by testing three samples with differing concentrations of CD25/IL-2R alpha in ELISA kits from four different production batches/lots.
| Lots | Lot 1 (pg/ml) | Lot 2 (pg/ml) | Lot 3 (pg/ml) | Lot 4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 140 | 116 | 139 | 115 | 127 | 12.01 | 9.4% |
| Sample 2 | 719 | 735 | 616 | 709 | 694 | 46.4 | 6.6% |
| Sample 3 | 2353 | 2484 | 2499 | 2400 | 2434 | 60.08 | 2.4% |
Protein Target Info & Infographic
Gene/Protein Information For IL2RA (Source: Uniprot.Org, NCBI)
Gene Name
IL2RA
Full Name
Interleukin-2 receptor subunit alpha
Weight
30819 MW
Alternative Names
Interleukin-2 receptor subunit alpha;IL-2 receptor subunit alpha;IL-2-RA;IL-2R subunit alpha;IL2-RA;TAC antigen;p55;CD25;IL2RA; IL2RA CD25, IDDM10, IL2R, IMD41, TCGFR, p55 interleukin 2 receptor subunit alpha interleukin-2 receptor subunit alpha|IL-2 receptor subunit alpha|IL-2R subunit alpha|TAC |interleukin 2 receptor, alpha
*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".For more info on IL2RA, check out the IL2RA Infographic
We have 30,000+ of these available, one for each gene! check them out.
In this infographic you will see the following information for IL2RA: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].
Specific Publications For Human CD25/IL-2sR Alpha ELISA Kit PicoKine® (EK0400)
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Customer Q&As
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7 Customer Q&As for Human CD25/IL-2sR Alpha ELISA Kit PicoKine®
Question
Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for quantification CD25 using the EK0400 Human CD25 Picokine® ELISA Kit?
Verified Customer
Verified customer
Asked: 2019-04-04
Answer
A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may alter the stabilityof samples. It is recommend to assay sample on-stat collection when possible, or aliquot into single use volumes and store samples frozen. limit repetitive freeze-thaw cycles with the stored samples to avoid protein degradation.
Boster Scientific Support
Answered: 2019-04-04
Question
Q: how are cell lysates prepared for use in Picokine® ELISA kits?
S. Wilson
Verified customer
Asked: 2018-12-10
Answer
A: in those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer may impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.
Boster Scientific Support
Answered: 2018-12-10
Question
Q: how to analyze ELISA data? I measured CD25 level in cell lysates.
Verified Customer
Verified customer
Asked: 2018-05-13
Answer
A: we recommend you this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online
Boster Scientific Support
Answered: 2018-05-13
Question
How should I reconstitute this lyophilized standard (Control for Human CD25/IL-2sR Alpha ELISA Kit)? For QC purpose, what is the mean ± 2*standard deviation value for this control?
Verified Customer
Verified customer
Asked: 2017-05-26
Answer
Add 1 mL of the diluent buffer to get a 500 pg/mL solution. The mean ± 2*standard deviation for this control is 486 ± 50 pg.
Boster Scientific Support
Answered: 2017-05-26
Question
Q: What is the optimal O.D. value for CD25 ELISA kit? I used your CD25 ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. can I consider these samples contain CD25 even though the O.D. values are not very high?
L. Tiwari
Verified customer
Asked: 2017-04-24
Answer
A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. what you should focus on is whether your sample O.D. values are statistically significantly higher than your blank values. regarding your protocol, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. typically, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by converting cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this CD25 ELISA kit to have sensitivity of 5pg/ml, that means the minimum amount of CD25 that can be declared/interpreted as positive by the above standard is 5pg/ml.
Boster Scientific Support
Answered: 2017-04-24
Question
Q: is it okay to use citrate plasma as samples in Human CD25 Picokine® ELISA Kit (Catalog # EK0400)?
V. Thomas
Verified customer
Asked: 2016-08-22
Answer
A: Chelating agents such as EDTA, Heparin and Citrate can sequester metal ions from the functional domain of CD25 causing disruption of its protein structure. CD25 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes required for detection and block the antigen antibody reaction. We have tested the CD25 ELISA, treating samples with a number of anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.
Boster Scientific Support
Answered: 2016-08-22
Question
Q: if the enzyme conjugated CD25 antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the CD25 antigen?
Verified Customer
Verified customer
Asked: 2015-11-15
Answer
A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.
Boster Scientific Support
Answered: 2015-11-15
