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Cynomolgus IFN-Beta ELISA Kit (Serum, Plasma, TCM)

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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.

$755.00

Pack Size
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%

Intra-Assay ≤ 8%

 

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: 

 

  1. Krause et al. (2005). Pharma Ther. 106 (3):299-346.

  2. Weinstock-Guttman et al. (2008). Expert Opin Biol Ther. 8(9):1435-47.

  3. Lee et al. (2007). Mol Cells. 23(1):1-10. 

  4. Heim. (1999). J Recept Signal Transduct Res. 19(1-4):75-120.

  5. Taniguchi et al . (2002). Curr Opin Immunol. 14(1):111-6.

  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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

Documentation

Certificate of Analysis (CoA), Protocol, and Safety Data Sheet (SDS)
46415-1 CoA & Protocol (One-Page)

46415-1 Certificate of Analysis (CoA) & Protocol (One-Page)

46415 Protocol (Full)

46415 Protocol (Full)

46415-1 CoA

46415-1 Certificate of Analysis (CoA)

46415 SDS

46415 Safety Data Sheet

46415 Product Flyer

46415 Product Flyer