Description
Human IL-28B ELISA Kit
The Human IL28B (Interleukin 28B) ELISA Kit is specifically designed to accurately measure IL28B levels in human biological samples such as serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures precise and reliable results for a variety of research applications.Interleukin 28B is a key cytokine that plays a crucial role in the immune response to viral infections, particularly hepatitis C.
Its levels are often altered in individuals with chronic viral infections, making it a valuable biomarker for studying immune responses and developing targeted therapies.This ELISA kit provides researchers with the tools needed to study the role of IL28B in viral infections and other immune-related conditions, ultimately contributing to the advancement of treatment strategies and personalized medicine.
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 15.63-1000 pg/mL |
| Sensitivity: | 9.38 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human IL28B ELISA Kit in samples. No significant cross-reactivity or interference between Human IL28B ELISA Kit and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human IL28B. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IL28B 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 IL28B, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human IL28B. The concentration of Human IL28B in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | IL-28B: Cytokine with immunomodulatory activity. Up-regulates MHC class I antigen expression. Displays potent antiviral activity. Also displays antitumor activity. Ligand for the heterodimeric class II cytokine receptor composed of IL10RB and IL28RA. The ligand/receptor complex seems to signal through the Jak-STAT pathway. Belongs to the IL-28/IL-29 family. |
| UniProt Protein Details: | Protein type:Secreted, signal peptide; Secreted Chromosomal Location of Human Ortholog: 19q13. 13 Cellular Component: extracellular space Molecular Function:cytokine activity Biological Process: positive regulation of immune response; negative regulation of viral genome replication; JAK-STAT cascade; defense response to virus Disease: Hepatitis C Virus, Susceptibility To |
| NCBI Summary: | This gene encodes a cytokine distantly related to type I interferons and the IL-10 family. This gene, interleukin 28A (IL28A), and interleukin 29 (IL29) are three closely related cytokine genes that form a cytokine gene cluster on a chromosomal region mapped to 19q13. Expression of the cytokines encoded by the three genes can be induced by viral infection. All three cytokines have been shown to interact with a heterodimeric class II cytokine receptor that consists of interleukin 10 receptor, beta (IL10RB) and interleukin 28 receptor, alpha (IL28RA). [provided by RefSeq, Jul 2008] |
| UniProt Code: | Q8IZI9 |
| NCBI GenInfo Identifier: | 300669648 |
| NCBI Gene ID: | 282617 |
| NCBI Accession: | Q8IZI9. 2 |
| UniProt Secondary Accession: | Q8IZI9,Q6VN56, Q7Z4J3, Q8IWL6, A2BDE1, |
| UniProt Related Accession: | Q8IZI9 |
| Molecular Weight: | 196 |
| NCBI Full Name: | Interferon lambda-3 |
| NCBI Synonym Full Names: | interferon, lambda 3 |
| NCBI Official Symbol: | IFNL3 |
| NCBI Official Synonym Symbols: | IL28B; IL28C; IL-28B |
| NCBI Protein Information: | interferon lambda-3; IL-28C; IFN-lambda-3; IFN-lambda-4; interleukin 28C; interleukin-28B; interleukin-28C; cytokine Zcyto22; interferon lambda-4; interferon, lambda 4; interleukin 28B (interferon, lambda 3) |
| UniProt Protein Name: | Interferon lambda-3 |
| UniProt Synonym Protein Names: | Cytokine Zcyto22; Interleukin-28B; IL-28B; Interleukin-28C; IL-28C |
| Protein Family: | Interferon |
| UniProt Gene Name: | IFNL3 |
| UniProt Entry Name: | IFNL3_HUMAN |
As the OD 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) | O.D | Average | Corrected |
| 1000 | 2.432 2.488 | 2.46 | 2.375 |
| 500 | 1.645 1.689 | 1.667 | 1.582 |
| 250 | 0.994 0.984 | 0.989 | 0.904 |
| 125 | 0.439 0.473 | 0.456 | 0.371 |
| 62.5 | 0.261 0.253 | 0.257 | 0.172 |
| 31.25 | 0.191 0.181 | 0.186 | 0.101 |
| 15.63 | 0.131 0.143 | 0.137 | 0.052 |
| 0 | 0.082 0.088 | 0.085 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IL28B ELISA 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 IL28B ELISA 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) | 54.62 | 143.96 | 434.76 | 49.45 | 134.17 | 441.58 |
| Standard deviation | 3.16 | 7.63 | 17.43 | 3.17 | 7.37 | 17.00 |
| C V (%) | 5.79 | 5.30 | 4.01 | 6.41 | 5.49 | 3.85 |
Recovery
The recovery of Human IL28B ELISA 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) | 91-104 | 96 |
| EDTA plasma (n=5) | 86-101 | 93 |
| Cell culture media (n=5) | 89-105 | 96 |
Linearity
Samples were spiked with high concentrations of Human IL28B ELISA 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 (%) | 87-100 | 97-112 | 95-113 |
| Average (%) | 93 | 105 | 103 | |
| 1:4 | Range (%) | 92-106 | 85-97 | 89-100 |
| Average (%) | 97 | 91 | 94 | |
| 1:8 | Range (%) | 89-104 | 85-98 | 85-101 |
| Average (%) | 96 | 91 | 92 | |
| 1:16 | Range (%) | 88-101 | 82-94 | 84-97 |
| Average (%) | 94 | 87 | 89 |
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 ELISA 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 | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| 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 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
