Rat IL-2SR alpha CLIA Kit (RTES00522)

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RTES00522
€649

Description

ELISA Kit Technical ManualMSDS

Rat IL2SR alpha (Soluble Interleukin-2 Receptor) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Rat IL2sR alpha/CD25 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat IL2sR alpha/CD25 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat IL2sR alpha/CD25, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Rat IL2sR alpha/CD25. The concentration of Rat IL2sR alpha/CD25 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityRat
Detection methodChemiluminescence
Detection range15.63-1000 pg/mL
Sensitivity9.38 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Rat IL2SR alpha CLIA Kit in samples. No significant cross-reactivity or interference between Rat IL2SR alpha CLIA Kit and analogues was observed.

Typical Data

As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(pg/mL)
RLU Average Corrected
100050091
57259
5367553650
50022293
22685
2248922464
25010991
9431
1021110186
1254759
5043
49014876
62.52646
2260
24532428
31.251353
1209
12811256
15.63677
739
708683
025
25
25--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat IL2SR alpha CLIA Kit were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat IL2SR alpha CLIA Kit were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)53.22145.79338.5956.57145.50325.57
Standard deviation6.0514.8425.024.9010.4235.88
C V (%)11.3710.187.398.667.1611.02

Recovery

The recovery of Rat IL2SR alpha CLIA Kit spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)88-10394
EDTA plasma (n=5)99-113106
Cell culture media (n=5)96-111101

Linearity

Samples were spiked with high concentrations of Rat IL2SR alpha CLIA Kit and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

    Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)93-10686-10098-117
Average (%)9892107
1:4Range (%)89-10599-11385-100
Average (%)9610792
1:8Range (%)86-9995-11295-109
Average (%)92102101
1:16Range (%)91-10289-10092-107
Average (%)979498

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro CLIA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Rat IL2SR alpha CLIA Kit (RTES00522) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Rat IL2SR alpha CLIA Kit (RTES00522) Protein Information

UniProt Protein Function:IL2RA: Receptor for interleukin-2. Non-covalent dimer of an alpha and a beta subunit. IL2R exists in 3 different forms: a high affinity dimer, an intermediate affinity monomer (beta subunit), and a low affinity monomer (alpha subunit). The high and intermediate affinity forms also associate with a gamma subunit.
UniProt Protein Details:

Protein type:Membrane protein, integral

Chromosomal Location of Human Ortholog: 10p15-p14

Cellular Component: integral to membrane; plasma membrane; external side of plasma membrane

Molecular Function:interleukin-2 receptor activity; drug binding; interleukin-2 binding

Biological Process: Notch signaling pathway; regulation of T cell homeostatic proliferation; apoptosis; negative regulation of immune response; negative regulation of defense response to virus; positive regulation of activated T cell proliferation; cell proliferation; negative regulation of T cell proliferation; inflammatory response to antigenic stimulus; cell surface receptor linked signal transduction; positive regulation of T cell differentiation; negative regulation of inflammatory response; immune response; inflammatory response; activated T cell apoptosis

Disease: Interleukin 2 Receptor, Alpha, Deficiency Of; Diabetes Mellitus, Insulin-dependent, 10

NCBI Summary:The interleukin 2 (IL2) receptor alpha (IL2RA) and beta (IL2RB) chains, together with the common gamma chain (IL2RG), constitute the high-affinity IL2 receptor. Homodimeric alpha chains (IL2RA) result in low-affinity receptor, while homodimeric beta (IL2RB) chains produce a medium-affinity receptor. Normally an integral-membrane protein, soluble IL2RA has been isolated and determined to result from extracellular proteolyisis. Alternately-spliced IL2RA mRNAs have been isolated, but the significance of each is presently unknown. Mutations in this gene are associated with interleukin 2 receptor alpha deficiency.[provided by RefSeq, Nov 2009]
UniProt Code:P26897
NCBI GenInfo Identifier:4557667
NCBI Gene ID:3559
NCBI Accession:NP_000408.1
UniProt Secondary Accession:P26897,P26897,
UniProt Related Accession:P01589
Molecular Weight:30,819 Da
NCBI Full Name:interleukin-2 receptor subunit alpha
NCBI Synonym Full Names:interleukin 2 receptor, alpha
NCBI Official Symbol:IL2RA  
NCBI Official Synonym Symbols:CD25; IL2R; TCGFR; IDDM10  
NCBI Protein Information:interleukin-2 receptor subunit alpha; p55; IL2-RA; IL-2-RA; TAC antigen; IL-2R subunit alpha; IL-2 receptor subunit alpha
UniProt Protein Name:Interleukin-2 receptor subunit alpha
UniProt Synonym Protein Names:TAC antigen; p55
UniProt Gene Name:IL2RA  
UniProt Entry Name:IL2RA_HUMAN
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Additional Information

Product type:
ELISA
Reactivity:
Rat
ELISA Type:
CLIA
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