Multiplex Assay Kit for Filaggrin (FLG) ,etc. by FLIA (Flow Luminescence Immunoassay)

Profilaggrin

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Filaggrin (FLG) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Filaggrin (FLG) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Filaggrin (FLG) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Filaggrin (FLG) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-105 101
EDTA plasma(n=5) 82-94 89
heparin plasma(n=5) 97-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Filaggrin (FLG) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Filaggrin (FLG) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Filaggrin (FLG) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 90-103% 94-102% 87-101% 81-91%
EDTA plasma(n=5) 92-105% 85-99% 80-93% 80-96%
heparin plasma(n=5) 99-105% 80-98% 93-101% 93-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:FLG) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Journal of the Egyptian Women's Dermatologic Society Filaggrin R501x and 2282del4 gene mutations and its tissue levels in patients with chronic plaque psoriasis LWW: 01000
?Inflammation &amp; Allergy-Drug Targets Neuromodulatory and Anti-Inflammatory Ingredient for Sensitive Skin: In Vitro Assessment Pubmed:24931812
PLoS One Protective Effects of Triphala on Dermal Fibroblasts and Human Keratinocytes Pubmed:26731545
Journal of food biochemistry In vitro anti-inflammatory and skin protective properties of extract comprising ceramides from de-oiled rice bran doi:10.1111
European Journal of Pharmacology Imiquimod-induced psoriasis-like inflammation in differentiated Human keratinocytes: Its evaluation using curcumin. pubmed:28736282
Journal of Dermatological Science A novel, topical, nonsteroidal, TRPV1 antagonist, PAC-14028 cream improves skin barrier function and exerts anti-inflammatory action through modulating epidermal … Pubmed:29752146
Journal of Traditional and Complementary Medicine In vitro anti-inflammatory and skin protective properties of Virgin coconut oil 10.1016:j.jtcme.2017.06.012
Clinical and Molecular Allergy Markers of atopic dermatitis, allergic rhinitis and bronchial asthma in pediatric patients: correlation with filaggrin, eosinophil major basic protein and …
Intractable & Rare Diseases Research Filaggrin, major basic protein and leukotriene B4: Biomarkers for adult patients of bronchial asthma, atopic dermatitis and allergic rhinitis Pubmed: 30560019
Dermatology and Therapy Epidermal and Dermal Hallmarks of Photoaging are Prevented by Treatment with Night Serum Containing Melatonin, Bakuchiol, and Ascorbyl …
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J Dermatol Filaggrin‐gene mutation has minimal effect on the disease severity in the lesions of atopic dermatitis 34322929
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RPJ103Hu02 Recombinant Filaggrin (FLG) Positive Control; Immunogen; SDS-PAGE; WB.
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LAJ103Hu83 FITC-Linked Polyclonal Antibody to Filaggrin (FLG) WB; IHC; ICC; IF.
LAJ103Hu81 FITC-Linked Polyclonal Antibody to Filaggrin (FLG) WB; IHC; ICC; IF.
MAJ103Hu21 Monoclonal Antibody to Filaggrin (FLG) WB; IHC; ICC; IP.
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