Cynomolgus IFN-Beta ELISA Kit (Serum, Plasma, TCM)
Catalog Number: 46415
This ELISA quantifies Cynomolgus IFN-Beta in serum, plasma, and tissue culture media (TCM) with an LLOQ of 5.47 pg/ml. Recombinant mammalian Cynomolgus IFN-Beta is provided as the ELISA standard.
Product Name: VeriKine Cynomolgus Interferon-Beta ELISA Kit (Serum, Plasma, TCM)
$755.00
Product Info
| Matrix Compatibility | Serum, Plasma, Cell Culture Supernatant |
|---|---|
| Assay Range | 5.47 - 350 pg/ml |
| LLOQ |
5.47 pg/ml Need more sensitivity? Check out our Sample Testing Services |
| Assay Length | 3 hours |
| Specificity | Cynomolgus (Macaca fascicularis) IFN-Beta |
The VeriKine Cynomolgus IFN-Beta ELISA has been developed to measure low/basal levels of non-human primate IFN-Beta (Macaca fascicularis IFN-Beta) in a variety of sample matrices including serum, plasma and tissue culture media (TCM) by sandwich enzyme-linked immunosorbent assay (ELISA).
Specifications
| CVs and Spike Recovery |
Inter-Assay ≤ 10%
Average Spike Recovery: 94% |
|---|---|
| Storage | 2-8°C |
| Expiration Date | One year from the date of manufacture |
| Shipping Condition | Wet Ice |
Materials Provided
- Pre-coated microtiter plate
- Plate Sealers
- Wash Solution Concentrate
- Cynomolgus Interferon Beta Standard, 100,000 pg/ml
- Standard Diluent
- Sample Buffer
- Antibody Concentrate
- HRP Conjugate Concentrate
- Assay Diluent
- TMB Substrate Solution
- Stop Solution
Additional Materials Required (Not Provided)
- Microplate reader capable of reading an OD at a wavelength of 450 nm
- Variable volume microtiter pipettes
- Adjustable multichannel pipette (50-300 μl)
- Reagent reservoirs
- Wash bottle or plate washing system
- Distilled or deionized water
- Serological pipettes (1, 5, 10 or 25 ml)
- Disposable pipette tips (polypropylene)
- Plate shaker
Tech Info & Data
Intra and Inter-Assay CVs to measure Precision |
|||
| L | M | H | |
| Concentration (pg/ml) | 6 | 40 | 200 |
| Intra-Assay CV (%) | 2.5 | 1.6 | 2.8 |
|
Inter-Assay CV (%) Operator 1 (n=4) |
5.4 | 4.3 | 2.3 |
|
Inter-Assay CV (%) Operator 2 (n=3) |
4.4 | 3.4 | 5.8 |
| Average % Recovery | 93.7 | 97.1 | 92.4 |
Background
Interferons (IFNs) are a group of cytokines that exhibit pleiotropic activities that play major roles in both innate and adaptive immunity. Type I IFNs consist of multiple IFN Alpha (α) genes and at least one IFN-Beta (β) gene in most vertebrates. IFN-β is used therapeutically to treat Multiple Sclerosis. Although well studied in humans and mice, the role of IFN-Beta in Rhesus and Cynomolgus monkey responses is relatively limited.
IFN-β expression and secretion are primarily induced by signaling from pattern recognition receptors such as Toll-like (TLR) and RIG-I-like receptors (RLR). Overall, IFN-β is part of the first wave of cytokine response in cells. Pathogen infection can result in the activation of interferon regulatory factor 3 (IRF3) which functions in trans to activate IFN-β gene transcription.
Following expression and secretion, IFN-β binds to a transmembrane heterodimeric receptor chain consisting of IFNAR1 & IFNAR2 on infected (autocrine) or neighboring cell (paracrine) surfaces. Receptor binding promotes a signal transduction cascade consisting of components of the JAK-STAT signaling pathway. This results in the expression of many genes including interferon regulatory factor 7 (IRF7) which upregulates the expression of many IFN-α subtype proteins. The IRF3/IRF7 signaling cascade is important for the initial and progressive responses to pathogens wherein hundreds of genes are regulated in a coordinated, temporal manner.
IFN-β is biologically unique when compared to other interferons. Studies have shown that IFN-β has overlapping and distinct gene expression patterns as compared to IFN-α. It appears that IFN-β binds to the Type 1 IFN receptor with higher affinity than other Type I IFNs and that it may also regulate receptor internalization in a different manner.
Basal levels of Type I IFNs, including IFN-β, are not fully understood. They are believed to be important for a robust response to pathogens and may play additional roles in cellular homeostasis.
This immunoassay has been developed to measure low levels of non-human primate IFN-Beta in a variety of sample matrices by sandwich enzyme-linked immunosorbent assay (ELISA). Interferon binds to plates coated with antibody and detection is accomplished using a biotinylated detection antibody followed by streptavidin conjugated to horseradish peroxidase (HRP). The substrate is tetramethylbenzidine (TMB).
Citations
References:
- Krause et al. (2005). Pharma Ther. 106 (3):299-346.
- Weinstock-Guttman et al. (2008). Expert Opin Biol Ther. 8(9):1435-47.
- Lee et al. (2007). Mol Cells. 23(1):1-10.
- Heim. (1999). J Recept Signal Transduct Res. 19(1-4):75-120.
- Taniguchi et al . (2002). Curr Opin Immunol. 14(1):111-6.
- Lewerenz M, Mogensen KE, Uzé G. Shared receptor components but distinct complexes for alpha and beta interferons. J Mol Biol. 1998 Sep 25;282(3):585-99.
- Jaks E, Gavutis M, Uzé G, Martal J, Piehler J. Differential receptor subunit affinities of type I interferons govern differential signal activation. J Mol Biol. 2007 Feb 16;366 (2):525- 39.
- Marijanovic Z, Ragimbeau J, van der Heyden J, Uzé G, Pellegrini S. Comparable potency of IFNalpha2 and IFN beta on immediate JAK/ST A T activation but differential downregulation of IFNAR2. Biochem J. 2007 Oct 1;407(1):141-51.
- Taniguchi T , T akaoka A. A weak signal for strong responses: interferon-alpha/beta revisited. Nat Rev Mol Cell Biol. 2001 May;2(5):378-86.
- Aziz N, Nishanian P, Mitsuyasu R, Detels R, Fahey JL. Variables That Affect Assays for Plasma Cytokines and Soluble Activation Markers. Clin Diagn Lab Immunol. 1999 Jan: (6)1:89-95.