Log In Sign up
The largest database of trusted experimental protocols

Phadia 100

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in Sweden, Italy

The Phadia 100 is a laboratory instrument designed for the detection and quantification of specific antibodies in patient samples. It utilizes fluoroenzyme immunoassay (FEIA) technology to measure levels of IgE, IgG, and other antibodies. The Phadia 100 provides accurate and reliable results for allergy and autoimmune testing applications.

Automatically generated - may contain errors

Lab products found in correlation

Phadia 250, by Thermo Fisher Scientific (3 mentions) Immunocap isac, by Thermo Fisher Scientific (1 mentions) Human il4 il10 enzyme immunoassay kit, by Boster Bio (1 mentions) Human elisa assay kit, by BioVendor (1 mentions) Cobas c501 analyser, by Roche (1 mentions) Zeba desalt spin column, by Thermo Fisher Scientific (1 mentions) Quant tag biotin kit, by Vector Laboratories (1 mentions) Streptavidin immunocaps, by Thermo Fisher Scientific (1 mentions) Aluminum hydroxide gel, by Merck Group (1 mentions) Nab protein g spin columns, by Thermo Fisher Scientific (1 mentions) Macrosep advance centrifugal device, by Pall Corporation (1 mentions) Elia calprotectin, by Thermo Fisher Scientific (1 mentions) Immunocap, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

ImmunoCAP Phadia® 100 Laboratory System

18 protocols using phadia 100

1

Biotinylation and Immobilization of Recombinant Allergen

Check if the same lab product or an alternative is used in the 5 most similar protocols
For custom ImmunoCAP, recombinant allergen was dialyzed against 1 L of 0.1 M NaHCO3, 1 M NaCl buffer. After dialysis, the protein concentration was measured. Biotin stock 10 mg/mL was used in 5-fold molar excess to bind the recombinant allergen (3 h, RT, mixed), followed by PBS dialysis and another measurement of the protein concentration. The biotinylated allergen was bound on Streptavidin ImmunoCAP o212 (Thermo Fisher Scientific) using a custom program (prewashed, loaded 50 µL of 100 µg/mL allergen/CAP, incubated 30’, washed) on a Thermo Scientific™ Phadia100, then detected with patient sera in a Thermo Scientific™ Phadia250 instrument according to manufacturer protocols.
Tamaș T.P., Buzan M.R., Zbîrcea L.E., Cotarcă M.D., Grijincu M., Păunescu V., Panaitescu C, & Chen K.W. (2023). Ragweed Major Allergen Amb a 11 Recombinant Production and Clinical Implications. Biomolecules, 13(1), 182.
+ Open protocol
+ Expand
2

Multiplex Measurement of Specific IgE Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total serum IgE levels were measured using Phadia 100; results were reported in kIU/L. Specific serum IgE antibodies were measured with the ImmunoCAP ISAC (Thermo Fisher Scientific, Waltham, MA USA). ImmunoCAP ISAC is a miniaturized immunoassay platform that allows for multiplex measurement of specific IgE antibodies to many allergen components using 20 μL of serum or plasma. Allergens are immobilized on a microarray chip to allow simultaneous measurement of specific IgE antibodies to 112 components from 51 allergen sources.21 (link) Test results were measured with a biochip scanner and results were reported in ISAC standardized units (ISU) and categorized based on the manufacturer’s cutoff levels (< 0.3 ISU, undetectable or very low; 0.3-0.9 ISU, low; 1-14.9 ISU, moderate/high; and ≥ 15 ISU, very high). Values above 1 ISU were considered positive.
Siah K.T., Santosa A., Cheung C.K., Soh A.Y, & Bigliardi P.L. (2020). Atopic Patients Who Fulfilled Rome III Criteria for Irritable Bowel Syndrome Had Higher Animal Danders Sensitization. Journal of Neurogastroenterology and Motility, 26(2), 267-273.
+ Open protocol
+ Expand
3

Recombinant Allergen IgE Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
Specific IgE levels to recombinant proteins were determined by ImmunoCAP in Phadia 100 and 250 instruments (Thermo Fisher Scientific, Uppsala, Sweden) according to the manufacturer's instructions. In the case of Bet v 1, Api g 1.01 and Cor a 1.04, commercial ImmunoCAP tests were used, whereas for Pru av 1, Dau c 1.01, NCS and its variants, experimental ImmunoCAPs were manufactured by coupling the recombinant allergens individually to the solid phase, as described elsewhere [27] , [28] (link).
Berkner H., Seutter von Loetzen C., Hartl M., Randow S., Gubesch M., Vogel L., Husslik F., Reuter A., Lidholm J., Ballmer-Weber B., Vieths S., Rösch P, & Schiller D. (2014). Enlarging the Toolbox for Allergen Epitope Definition with an Allergen-Type Model Protein. PLoS ONE, 9(10), e111691.
+ Open protocol
+ Expand
4

Cow's Milk Allergy Biomarkers Assessment

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total IgE and specific IgE against of cow’s milk proteins (nBos d4; nBos d5; nBos d6; nBos d8) were assessed by enzymatic immunoassay (Phadia 100 ThermoFisher Scientific CAP system, Rodano Milano, Italy). Measurements were expressed as kilounits per liter (kU/L).
The concentrations of IL-4 and IL-10 were measured with a Human IL4/IL10 Enzyme immunoassay kit (Boster Biological Technology, Ltd., Fremont, CA, USA). The IL-5 and IFN- concentrations were measured using the human ELISA assay kit (BioVendor, Brno, Czech Republic). The minimum detection concentrations were 15.6 pg/mL for IL-4, 7.8 pg/mL for IL-5 and IL-10, and 0.78 pg/mL for IFN-γ.
Paparo L., Nocerino R., Bruno C., Di Scala C., Cosenza L., Bedogni G., Di Costanzo M., Mennini M., D’Argenio V., Salvatore F, & Berni Canani R. (2019). Randomized controlled trial on the influence of dietary intervention on epigenetic mechanisms in children with cow’s milk allergy: the EPICMA study. Scientific Reports, 9, 2828.
+ Open protocol
+ Expand
5

Coconut Milk IgE Reactivity Evaluation

Check if the same lab product or an alternative is used in the 5 most similar protocols
IgE reactivity to coconut milk was evaluated using Phadia ImmunoCAP (f36) (Thermo Fisher Scientific, Uppsala, Sweden) using the Phadia 100 (Table 1). The test was performed according to the manufacturer’s instructions. The cutoff value was set as > 0.35 kUA/L for sIgE positivity.
Iddagoda J., Gunasekara P., Handunnetti S., Jeewandara C., Karunatilake C., Malavige G.N., de Silva R, & Dasanayake D. (2022). Identification of allergens in coconut milk and oil with patients sensitized to coconut milk in Sri Lanka. Clinical and Molecular Allergy : CMA, 20, 14.
+ Open protocol
+ Expand
6

Fecal Calprotectin Measurement Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols
Based on study protocol (Figure 1), extracts were analysed with fluorescence enzyme immunoassay (FEIA), EliATM Calprotectin on Phadia 100 analyser (Thermo Fisher, Uppsala, Sweden) and/or PETIA, Bühlmann fCAL® Turbo on Roche Cobas c501 analyser (Roche diagnostics, Manheim, Germany). Measuring ranges for FEIA and PETIA are 15 - 3000 mg/kg (within-laboratory coefficient of variation, CV = 4.8%) and 20 - 2000 mg/kg (within-laboratory CV = 4.9%), respectively. Cut-off value for both methods is set at 50 mg/kg (negative). Values 50 - 200 mg/kg represent the “grey zone” and > 200 mg/kg active inflammation in gastrointestinal tract (14 , 22 ).
Juricic G., Brencic T., Tesija-Kuna A., Njegovan M, & Honovic L. (2019). Faecal calprotectin determination: impact of preanalytical sample treatment and stool consistency on within- and between-method variability. Biochemia Medica, 29(1), 010707.
+ Open protocol
+ Expand
7

IgE Antibody Binding Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Direct IgE antibody binding assays were performed by ImmunoCAP. Biotinylation of
the allergens was carried out using the EZ-Link Sulfo-NHS-LC-Biotin, No-Weigh™
Format (Thermo Fisher Scientific Inc., Pierce, Rockford, IL), with biotin added at a
10-fold molar excess to the allergens. Zeba™ Desalt Spin Columns (Thermo Fisher
Scientific Inc., Pierce, Rockford, IL) were used to remove any excess biotin from the
biotinylated allergens. A Quant*Tag Biotin Kit™ (Vector Laboratories, Inc.,
Burlingame, CA) was used to quantify the number of biotins per molecule. IgE detection and
quantification in plasma samples was determined using Specific IgE detection
streptavidin-ImmunoCAP on the Phadia® 100 (Thermo Fisher Scientific, Portage, MI).
After testing several concentrations of allergen (from 0.25 to 10 µg/ml),
streptavidin ImmunoCAPs were loaded with 5 µg/CAP of biotinylated allergen and
incubated for 30 minutes. IgE measurements were performed according to standard ImmunoCAP
procedures.
Glesner J., Vailes L.D., Schlachter C., Mank N., Minor W., Osinski T., Chruszcz M., Chapman M.D, & Pomés A. (2016). Antigenic determinants of Der p 1: Specificity and cross-reactivity associated with IgE antibody recognition. Journal of immunology (Baltimore, Md. : 1950), 198(3), 1334-1344.
+ Open protocol
+ Expand
8

Optimizing Allergen-Loaded Immunocaps

Check if the same lab product or an alternative is used in the 5 most similar protocols
Streptavidin ImmunoCAPs (Thermo Fisher Scientific, Portage, MI) were loaded and incubated on a Phadia 100, with the biotinylated allergen at amounts ranging from 0.5 to 10 µg/CAP. Two different human plasma samples from individuals allergic to the allergen (that had been originally tested for IgE binding to 2–3 µg/CAP) were selected for optimization experiments. Their IgE levels to the allergen-loaded CAPs were measured in a Phadia 250 following manufacturer’s instructions (Thermo Fisher Scientific, Portage, MI) to select optimal amount of biotinylated allergen to be loaded to the Streptavidin ImmunoCAPs.
Pomés A., Schulten V., Glesner J., da Silva Antunes R., Sutherland A., Bacharier L.B., Beigelman A., Busse P., Frazier A, & Sette A. (2021). IgE and T Cell Reactivity to a Comprehensive Panel of Cockroach Allergens in Relation to Disease. Frontiers in Immunology, 11, 621700.
+ Open protocol
+ Expand
9

Measuring Serum IgE and WBC Counts

Check if the same lab product or an alternative is used in the 5 most similar protocols
Serum total immunoglobulin E (IgE) was measured in our research lab from plasma in duplicate on a Phadia 100 detection system (ThermoFisher Scientific, Uppsala, Sweden). If both measurements were not within 10% concordance, a 3rd measurement was assayed. White blood cell (WBC) counts were obtained from complete blood counts with differentials using commercially available and Clinical Laboratory Improvement Amendments (CLIA)-certified laboratories through Quest Diagnostics in the GALA II dataset and through UCSF Clinical Laboratories for SAGE. Serum total IgE and WBC counts were specified as continuous predictors. However, the distributions of serum total IgE, basophils, and eosinophils were highly right-skewed and were log-transformed for analysis purposes.
Wohlford E.M., Huang P.F., Elhawary J.R., Millette L.A., Contreras M.G., Witonsky J., Holweg C.T., Oh S.S., Lee C., Merenda C., Rabin R.L., Araojo R., Mak A.C., Eng C.S., Hu D., Huntsman S., LeNoir M.A., Rodríguez-Santana J.R., Borrell L.N, & Burchard E.G. (2021). Racial/Ethnic differences in eligibility for asthma biologics among pediatric populations. The Journal of allergy and clinical immunology, 148(5), 1324-1331.e12.
+ Open protocol
+ Expand
10

Murine OVA-specific IgG Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Murine OVA-specific IgG was prepared from pooled sera from mice immunized i.p. with alum (50μg protein precipitated to 1mg aluminum hydroxide gel (Sigma Aldrich, St. Louis, MO) on days 0 and 7, followed by 10μg protein on days 14 and 21). Control IgG was prepared from mock-immunized mice. Human IgG was prepared from pooled sera samples from patients who had undergone oral immunotherapy for peanut allergy or from non-allergic donors. IgG was isolated over Nab protein G spin columns (Thermo Scientific, Waltham, MA), concentrated and dialyzed with Macrosep Advance Centrifugal Devices carrying a 100 kDa cut-off (Pall Corporation, Westborough, MA) and filter-sterilized with 0.2μM syringe filters (Millex, EMD Millipore, Billerica, MA). Murine OVA-specific IgG concentrations were determined by ELISA, and mice were injected with 100μg IgG1. Peanut-specific IgG4 in human IgG preparations was quantified by ImmunoCAP on a Phadia 100 (ThermoScientific, Kalamazoo, MI).
Burton O.T., Tamayo J.M., Stranks A.J., Koleoglou K.J, & Oettgen H.C. (2017). Allergen-specific IgG antibodies signaling via FcγRIIb promote food tolerance. The Journal of allergy and clinical immunology, 141(1), 189-201.e3.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!