Representative Human IFN-Beta Standard Curves in Various Matrices (see Technical Data Sheet)

Matrix Compliance of Standard Diluent and Three Different TCMs (see Technical Data Sheet)

Parallelism of Endogenous Samples (see Technical Data Sheet)

Sensitivity in Stimulated PBMC Samples (see Technical Data Sheet)

Endogenous Levels of IFN-Beta Quantified in Sendai Virus-Induced HEK293 Cells (see Technical Data Sheet)

Representative Standard Curves comparison

sIFNAR2 Receptor Effect in PBL’s 41435 ELISA (see Technical Data Sheet)

sIFNAR2 Receptor Interference (see Technical Data Sheet)

Human IFN-Beta TCM ELISA, High Sensitivity

Catalog Number: 41435

This ELISA quantitates the amount of IFN-Beta in tissue culture media (TCM) with an LLOQ of 2.34 pg/ml. This kit provides accurate IFN-Beta quantification without interference from soluble IFN receptor (sIFNAR2).

Product Name: VeriKine-HS Human Interferon-Beta TCM ELISA Kit

$545.00

$645.00

Matrix Compatibility	Tissue Culture Media (TCM) only
Assay Range	2.34 - 150 pg/ml
LLOQ	2.34 pg/ml Need more sensitivity? Check out our Sample Testing Services
Assay Length	3 hours
Specificity	Human IFN-β

The VeriKine-HS Human IFN-Beta TCM ELISA Kit is designed for accurate quantification of low-level human interferon beta in buffers or tissue culture media (TCM) by sandwich ELISA. The LLOQ of this kit is 2.34 pg/ml.

Note: This kit is not intended for use with human serum or plasma samples due to a significantly elevated risk of false-positive results.

*For complex biological matrices such as plasma, healthy sera, and disease sera, we recommend our Human IFN-Beta Serum ELISA Kit, High Sensitivity (Cat. No. 41415-1).

CVs	Inter-Assay < 8% Intra-Assay < 10%
Cross-reactivity	No cross-reactivity detected with: Human IFN-α, IFN-γ, IFN-ω, IL-6 Mouse IFN-α, IFN-β Rat IFN-β
Synonyms	Human Beta Interferon, Human Fibroblast IFN, Human Type I Interferon Beta, Human IFNb
Storage	2-8ºC
Expiration Date	12 months from the date of manufacture
Shipping Condition	Wet Ice

Materials Provided

Pre-coated microtiter plate
Plate Sealers
Wash Solution Concentrate
Human IFN Beta 1a Standard, 100,000 pg/ml
Standard Diluent
Sample Buffer
Antibody Concentrate
HRP Conjugate Concentrate
Assay Diluent
TMB Substrate
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

Representative Human IFN-Beta Standard Curves in Various Matrices

41435 Representative Standard Curves — Human IFN-Beta standard curves were prepared in Tissue Culture Media (TCM) and Standard Diluent (SD). Three different TCM were used: RPMI, DMEM and MEM. TCM were supplemented with 10% FBS. This figure shows the mean of two runs and error bars indicate the standard deviation.

Intra and Inter-Assay CVs to measure Precision
Matrix	Standard Diluent	TCM
n	7	18
Intra-Assay CV (%)	4.10	4.06
Inter-Assay CV (%)	2.33	3.16

Spike Recovery
TC Media Spike Sample	1	2	3
Target Conc. (pg/ml)	100	25	5
Mean Recovery (%)	101.1	95.7	91.4
Range (%)	97-106	90-102	85-96

Matrix Compliance for Standard Diluent and Three Different TCM

41435 Fig 2 — Human IFN-Beta standard curves were prepared in Tissue Culture Media (TCM) and Standard Diluent (SD). Three different TCM were used: RPMI, DMEM and MEM, each supplemented with 10% FBS. The figure below shows the recovery of Human IFN-Beta Standard at the different concentrations for the various matrices. The SD curve was used as reference for calculating the percent recovery of IFN-Beta in TCM from the expected value. The figure shows the means of two runs, and error bars indicate the standard deviation.

Parallelism of Five Endogenous Samples

41435 Parallelism of Endogenous Samples — Five endogenous samples were tested and assayed in duplicate. Samples were serially diluted in TCM to the 32-fold dilution, which still lies within standard curve range. Percent recoveries are 100 ± 20% of neat value except for the 32nd dilution of Sample 5 (S5) which falls just beneath the bottom standard curve point of 2.34 pg/ml. Error bars indicate the standard deviation.

Sensitivity in Stimulated PBMC Samples

41435 Induction by TLR 7-8 — PBMCs were isolated from a healthy individual’s buffy coat and stimulated with 10 ug/ml of the TLR-7/8 agonist, R-848, to induce detectable levels of IFN-Beta in RPMI. Four densities of PBMCs were used (1E6, 2E6, 3E6 and 4E6 cells per ml), and supernatants were collected 24 and 48 hr post stimulation. The 1E6 PBMC samples fall below the LLOQ (Lower Limit of Quantitation) of 2.34 pg/ml.

Endogenous Levels of IFN-Beta Quantified in Sendai Virus-Induced HEK293 Cells

Representative Standard Curves

sIFNAR2 Receptor Interference in Competitor A’s ELISA

sIFNAR2 Receptor Effect in PBL’s 41435 ELISA

Quantitation of Total IFN-Beta in the presence of sIFNAR2

Background

Interferon Beta (IFN-Beta, IFNb) is synthesized and secreted by fibroblasts and many other cell types in response to varied stimuli in Toll-like receptor (TLR) dependent and independent mechanisms. Pathogen exposure can result in the activation of transcription factors interferon regulatory factor 3 (IRF3) and NF-kB resulting in IFN-Beta production in a stimuli and cell-dependent manner. Conversely, IFN-Beta aberrant expression may be associated with inflammation and autoimmunity.

4 Citations:

Luganini, A., et al., (2023), "Mechanisms of antiviral activity of the new hDHODH inhibitor MEDS433 against respiratory syncytial virus replication", Antiviral Res., 105734, PMID: 37852322, DOI: 10.1016/j.antiviral.2023.105734 (link)

Barnard, T.R. et al., (2023), "Zika virus NS3 and NS5 proteins determine strain-dependent differences in dsRNA accumulation in a host cell type-dependent manner", J Gen Virol., 104(6):001855, PMID: 37289497, DOI: 10.1099/jgv.0.001855 (link)

Gori Savellini, G. et al., (2023), "SARS-COV-2 omicron sub-lineages differentially modulate interferon response in human lung epithelial cells", Virus Res., 332:199134, PMID: 37192725, DOI: 10.1016/j.virusres.2023.199134 (link)

Kwart, D. et al., (2022), "Cancer cell-derived type I interferons instruct tumor monocyte polarization", Cell Rep., 41(10):111769, PMID: 36476866, DOI:10.1016/j.celrep.2022.111769 (link)

Documentation

Certificate of Analysis (CoA), Protocol, Technical Data Sheet (TDS), and Safety Data Sheet (SDS)

41435 CoA and Protocol

41435 Certificate of Analysis and Protocol

41435 Technical Data Sheet

41435 TDS

41435 SDS

41435 Safety Data Sheet

Human IFN-Beta TCM ELISA, High Sensitivity