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Mouse IFN-Alpha All Subtype ELISA Kit, High Sensitivity (Serum, Plasma, TCM)

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Mouse IFN-Alpha All Subtype ELISA Kit, High Sensitivity (Serum, Plasma, TCM)

Catalog Number: 42115

This Mouse Interferon Alpha ELISA has an LLOQ of 2.38 pg/ml and can detect all 14 mouse IFN-Alpha subtypes in serum, plasma, and tissue culture media. This ELISA provides total IFN-Alpha readout in approximately 2 hours.

$695.00

Pack Size
Product Info

Matrix Compatibility Serum, Plasma, Tissue Culture Media
Assay Range 2.38 - 152 pg/ml
LLOQ

2.38 pg/ml

Need more sensitivity? Check out our Sample Testing Services

Assay Length 1 hour, 54 minutes
Specificity All 14 Mouse IFN-α subtypes
CVs Inter-Assay CV: ≤ 10%
Intra-Assay CV: ≤ 8%

 

The VeriKine-HS Mouse IFN-Alpha ELISA kit has been developed to quantitate low levels of IFN-Alpha in mouse serum, plasma and cell/tissue culture media (10% FBS) using a sandwich immunoassay. The kit contains an anti-mouse IFN-Alpha detection antibody conjugated to biotin and quantified by streptavidin horseradish peroxidase (HRP) using tetramethyl-benzidine (TMB) as a substrate.

 

The kit standard is mouse IFN-Alpha 4 protein. It is important to run the standard curve in the same matrix as one’s samples unless one has demonstrated that the matrix does not significantly affect the signal.

 

Specifications

CVs and Spike Recovery

Inter-Assay CV ≤ 10%


Intra-Assay CV ≤ 8%

 

Spike Recovery ≥ 80% in Serum

Cross-reactivity

Very weak cross-reactivity detected with:

  • Human IFN-Alpha14

 

No cross-reactivity with:

  • Mouse IFN-Beta, IFN-Gamma
  • Rat IFN-Alpha1, IFN-Alpha14
Synonyms IFN alpha, IFN-Alpha, Multisubtype Interferon Alpha, alpha interferon, Type I interferon, IFN-α
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
  • Mouse IFN-Alpha 4 Standard, 10,000 pg/ml
  • Sample Diluent
  • Antibody Concentrate
  • Antibody Diluent
  • HRP Conjugate Concentrate
  • HRP 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

 

Representative Mouse IFN-Alpha Standard Curve in Sample Diluent

 

 

Intra and Inter-Assay CVs to measure Precision

Concentration (pg/ml) Endogenous Level
Low Medium High
Intra-Assay CV (%) 4.4 3.3 2.8
Inter-Assay CV (%) 7.8 5.8 8.7

 

 

Spike Recovery

Dilution Average Concentration (pg/ml) % Linear Adjusted Dilution
Not Adjusted for Dilution Adjusted for Dilution
2 39.5 79.1 -
4 19.1 76.6 96.8
8 9.7 77.3 97.8
16 4.9 78.8 99.7
32 2.5 80.7 102.0
64 1.2 77.7 98.2
Avg - 78.4 98.9

 

 

Endogenous Mouse IFN-Alpha Quantified in Plasma or Serum From Three Mouse Strains Using PBL’s High Sensitivity Mouse IFN-Alpha All Subtype ELISA

Total endogenous levels of mouse IFN-Alpha were quantified in pooled plasma (blue, left) and individual sera (red and green, right) for three mouse strains: CD-1, BALB/c, and C57BL/6. Results from this experiment led to the recommendation to pre-screen serum, as a significant portion of samples contain quantifiable levels of endogenous interferon alpha.

 

Background

 

Interferons (IFNs) are an extensive set of cytokines exhibiting pleiotropic activities and playing major roles in both innate and adaptive immunity. The Type I IFNs genes consist of multiple IFN-Alpha genes, at least one IFN-Beta gene in most vertebrates, and a few other family members including limitin in the mouse and IFN-Omega and -Epsilon in mice and humans. In the mouse, fourteen (14) IFN-Alpha genes have been cloned which exhibit at least 75% identity at the protein level. In the therapeutic setting, IFN-Alpha is used to treat viral diseases and certain cancers and is therefore frequently studied in mouse and other animal models of these diseases. Several lines of evidence indicate that IFN-Alpha proteins may be involved in the development of specific autoimmune diseases and that treatment with IFN-Alpha blocking antibodies may comprise a useful therapeutic strategy.

 

The expression of interferon genes (and therefore, proteins) is regulated by a group of interferon regulatory factors, i.e., the IRF family of proteins. Cellular levels of IRF3 and IRF7 regulate the expression of individual IFN-Alpha subtypes with most IFN-Alpha subtype genes requiring IRF7 for expression. IRF7 protein is either constitutively expressed or inducible, depending on cell type. In certain mouse cell lines, IRF7 expression is induced in particular by at least IFN-Beta and IFN-Alpha4 which then leads to expression of the other IFN-Alpha subtype genes. Additionally, murine IFN-Beta may be requisite for fibroblast synthesis of IFN-α but may not be required for other cells’ synthesis of IFN-Alpha. The IRF3/IRF7 signaling cascade is a focal point for initial and progressive cellular responses to pathogens where hundreds of genes are subsequently regulated in a temporally coordinated cascade.

 

Plasmacytoid dendritic cells (PDCs) are highly efficient producers of IFN-Alpha, but a wide variety of cells can also produce lower levels of these protein subtypes. The particular IFN-Alpha subtype genes that are expressed appear to be somewhat cell and stimulus-specific and, based on the results published from several gene induction studies, the time courses of induction of the individual protein subtypes are expected to differ markedly.

Citations

6 Citations:

 

  1. Kang, T. G. et al., (2022), "Viral coinfection promotes tuberculosis immunopathogenesis by type I IFN signaling-dependent impediment of Th1 cell pulmonary influx", Nat. Commun. 13(1):3155, PMID: 35672321, DOI: 10.1038/s41467-022-30914-3 (link)

  2. Choi, J, H. et al., (2020), "DUSP11-mediated control of 5'-triphosphate RNA regulates RIG-I sensitivity", Genes & Development, 34:1, PMID: 33184222, DOI: 10.1101/gad.340604.120 (link)

  3. Tang, X, et al. (2020). Sustained IFN-I stimulation impairs MAIT cell responses to bacteria by inducing IL-10 during chronic HIV-1 infection. Science Advances, 13 pgs. PMID: 32128419. (link)

  4. Leylek, Rebecca, et al. (2019). Integrated Cross-Species Analysis Identifies a Conserved Transitional Dendritic Cell Population. Cell Reports, 39 pgs. PMID: 31825848. (link)

  5. Gale, Emily, et al. (2019). A Nanoparticle Platform for Improved Potency, Stability, and Adjuvanticity of Poly(I:C). Advanced Therapeutics, 6 pgs. PMID: no PMID. (link)

  6. Hickerson, Brady, et al. (2020). Type I interferon underlies severe disease associated with Junín virus infection in mice. eLife. PMID: no PMID. (link)

  7.  

 

References:

 

  1. Krause CD, Pestka S. (2005) “Evolution of the Class 2 cytokines and receptors, and discovery of new friends and relatives,” Pharmacol Ther. 106(3):299-346.

  2. Chevaliez, S, Pawlotsky, JM (2009); “Interferons and their use in persistent viral Infections”; in Handbook of Exp Pharmacol; Vol.189; 203-41.

  3. Antonelli, G. (2008); “Biological basis for a proper clinical application of alpha interferons”; in New Microbiologica.; Vol. 31; Issue 3; 305-318.

  4. Ascierto, PA, Kirkwood, JM (2008); “Adjuvant therapy of melanoma with interferon: lessons of the past decade”; in Journal of Transl Med; Vol. 27; 60:62.

  5. Ferrantini, M, Capone, I, Belardelli, F (2007) “Interferon alpha and cancer: mechanisms of action and new perspectives of clinical use” in Biochimie.; Vol. 89; Issues 6 and 7; 884-893.

  6. Fleischmann, WR Jr, Wu, TG (2005); “Development of an interferon-based cancer vaccine protocol: application to several types of murine cancers”; in Methods Mol Med.; Vol. 116; 151-66.

  7. Meyer, O (2009); “Interferons and autoimmune disorders” in Joint Bone Spine. Vol. 76; Issue 5; 464-73.

  8. Burdick, LM, Somani, N, Somani, AK (2009); “Type I IFNs and their role in the development of autoimmune diseases”; in Expert Opinion Drug Safety; Vol. 8; Issue 4; 459-72.

  9. Kötter, I, Hamuryudan, V, Oztürk ,ZE, Yazici, H (Jan 7, 2010); “Interferon therapy in rheumatic diseases: state-of-theart”; in Current Opinion Rheumatology.

  10. Erlandsson, L, Blumenthal, R, Eloranta, ML, Engel, H, Alm, G, Weiss, S, Leanderson, T (1998); “Interferon-beta is required for interferon-alpha production in mouse fibroblasts”; in Curr Biol.; Vol. 8; Issue 4;223-226.

  11. Taniguchi T, Takaoka A (2002); “The interferon-alpha/beta system in antiviral responses: a multimodal machinery of gene regulation by the IRF family of transcription factors”; in Current Opinions in Immunology; Vol. 14; Issue 1; 111-116.

  12. Jegalian AG, Facchetti F, Jaffe ES (2009); “Plasmacytoid dendritic cells: physiologic roles and pathologic states”; in Advances in Anat Pathol; Vol.16; Issue 6; 392-404.

  13. Fitzgerald-Bocarsly P, Dai J, Singh S (2008); “Plasmacytoid dendritic cells and type I IFN: 50 years of convergent history”; in Cytokine Growth Factor Rev; Vol. 19; Issue 1; 3-19.

  14. Li L, Sherry B (2010); “IFN-alpha expression and antiviral effects are subtype and cell type specific in the cardiac response to viral infection”; in Virology; Vol. 396; Issue 1; 59-68.

  15. Baig E, Fish EN (2008); “Distinct signature type I interferon responses are determined by the infecting virus and the target cell”; in Antivir Ther.; Vol. 13; Issue 3; 409-22.

  16. Delhaye S, Paul S, Blakqori G, Minet M, Weber F, Staeheli P, Michiels T (2006); “Neurons produce type I interferon during viral encephalitis”; in Proc Natl Acad Sci USA.; Vol. 103; Issue 20; 7835-7840.

Documentation

Protocol, Certificate of Analysis (CoA), Technical Data Sheet (TDS), Safety Data Sheet (SDS)
42115-1 CoA & Protocol (One-Page)

42115-1 Certificate of Analysis (CoA) & Protocol (One-Page)

42115-1 Technical Data Sheet

42115-1 Technical Data Sheet

42115 SDS

42115 Safety Data Sheet

42115-1 Protocol (Full)

42115-1 Protocol (Full)

42115-1 CoA

42115-1 Certificate of Analysis (CoA)

42115 Product Notice Antibody Stock Change

42115 Product Notice Antibody Stock Change