Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 12.50-800 pg/mL |
| Sensitivity: | 7.50 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human IL1R2 CLIA Kit in samples. No significant cross-reactivity or interference between Human IL1R2 CLIA Kit and analogues was observed. |
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 Human IL1R2. 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 Human IL1R2 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 Human IL1R2, 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 Human IL1R2. The concentration of Human IL1R2 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| UniProt Protein Function: | IL-1RB: Non-signaling receptor for IL1A, IL1B and IL1RN. Reduces IL1B activities. Serves as a decoy receptor by competetive binding to IL1B and preventing its binding to IL1R1. Also modulates cellular response through non-signaling association with IL1RAP after binding to IL1B. IL1R2 (membrane and secreted forms) preferentially binds IL1B and poorly IL1A and IL1RN. The secreted IL1R2 recruits secreted IL1RAP with high affinity; this complex formation may be the dominant mechanism for neutralization of IL1B by secreted/soluble receptors. Belongs to the interleukin-1 receptor family. 2 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Membrane protein, integral Chromosomal Location of Human Ortholog: 2q12 Cellular Component: plasma membrane Molecular Function:interleukin-1 receptor activity; protein binding Biological Process: immune response |
| NCBI Summary: | The protein encoded by this gene is a cytokine receptor that belongs to the interleukin 1 receptor family. This protein binds interleukin alpha (IL1A), interleukin beta (IL1B), and interleukin 1 receptor, type I(IL1R1/IL1RA), and acts as a decoy receptor that inhibits the activity of its ligands. Interleukin 4 (IL4) is reported to antagonize the activity of interleukin 1 by inducing the expression and release of this cytokine. This gene and three other genes form a cytokine receptor gene cluster on chromosome 2q12. Alternative splicing results in multiple transcript variants and protein isoforms. Alternative splicing produces both membrane-bound and soluble proteins. A soluble protein is also produced by proteolytic cleavage. [provided by RefSeq, May 2012] |
| UniProt Code: | P27930 |
| NCBI GenInfo Identifier: | 124310 |
| NCBI Gene ID: | 7850 |
| NCBI Accession: | P27930. 1 |
| UniProt Secondary Accession: | P27930,Q6LCE6, Q9UE68, D3DVJ5, |
| UniProt Related Accession: | P27930 |
| Molecular Weight: | 33,622 Da |
| NCBI Full Name: | Interleukin-1 receptor type 2 |
| NCBI Synonym Full Names: | interleukin 1 receptor type 2 |
| NCBI Official Symbol: | IL1R2 |
| NCBI Official Synonym Symbols: | IL1RB; CD121b; IL1R2c; CDw121b; IL-1R-2; IL-1RT2; IL-1RT-2 |
| NCBI Protein Information: | interleukin-1 receptor type 2 |
| UniProt Protein Name: | Interleukin-1 receptor type 2 |
| UniProt Synonym Protein Names: | CD121 antigen-like family member B; CDw121b; IL-1 type II receptor; Interleukin-1 receptor beta; IL-1R-beta |
| Protein Family: | Interleukin-1 receptor |
| UniProt Gene Name: | IL1R2 |
| UniProt Entry Name: | IL1R2_HUMAN |
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 |
| 800 | 48756 56528 | 52642 | 52616 |
| 400 | 20690 22778 | 21734 | 21708 |
| 200 | 9785 9631 | 9708 | 9682 |
| 100 | 4434 4670 | 4552 | 4526 |
| 50 | 2252 2124 | 2188 | 2162 |
| 25 | 1133 987 | 1060 | 1034 |
| 12.50 | 504 514 | 509 | 483 |
| 0 | 25 27 | 26 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IL1R2 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 Human IL1R2 CLIA Kit were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 37.51 | 100.85 | 343.93 | 35.46 | 100.29 | 361.66 |
| Standard deviation | 3.04 | 9.33 | 32.85 | 4.32 | 10.38 | 29.48 |
| C V (%) | 8.10 | 9.25 | 9.55 | 12.18 | 10.35 | 8.15 |
Recovery
The recovery of Human IL1R2 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) | 94-111 | 101 |
| EDTA plasma (n=5) | 85-96 | 91 |
| Cell culture media (n=5) | 97-115 | 105 |
Linearity
Samples were spiked with high concentrations of Human IL1R2 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) | ||
| 1:2 | Range (%) | 92-103 | 91-106 | 87-97 |
| Average (%) | 97 | 97 | 92 | |
| 1:4 | Range (%) | 91-106 | 89-101 | 93-109 |
| Average (%) | 96 | 96 | 100 | |
| 1:8 | Range (%) | 98-113 | 84-99 | 102-116 |
| Average (%) | 105 | 91 | 110 | |
| 1:16 | Range (%) | 104-115 | 95-112 | 85-96 |
| Average (%) | 109 | 103 | 91 |
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 ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- 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.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- 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.
- 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.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- 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.
- Determine the RLU value of each well immediately.
