ELISA Kit for Fatty Acid Binding Protein 5 (FABP5)

E-FABP; EFABP; PA-FABP; PAFABP; Fatty Acid Binding Protein 5, Epidermal; Psoriasis-Associated; Epidermal-type fatty acid-binding protein; Psoriasis-associated fatty acid-binding protein homolog

Specificity

This assay has high sensitivity and excellent specificity for detection of Fatty Acid Binding Protein 5 (FABP5).
No significant cross-reactivity or interference between Fatty Acid Binding Protein 5 (FABP5) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Fatty Acid Binding Protein 5 (FABP5) and the recovery rates were calculated by comparing the measured value to the expected amount of Fatty Acid Binding Protein 5 (FABP5) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 79-101 83
heparin plasma(n=5) 96-103 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fatty Acid Binding Protein 5 (FABP5) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Fatty Acid Binding Protein 5 (FABP5) 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 Fatty Acid Binding Protein 5 (FABP5) 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) 98-105% 79-99% 80-97% 92-105%
EDTA plasma(n=5) 86-95% 87-101% 80-89% 84-96%
heparin plasma(n=5) 98-105% 85-101% 79-92% 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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Journal of Proteome Research Profiling Protein Markers Associated with Lymph Node Metastasis in Prostate Cancer by DIGE-based Proteomics Analysis PubMed: 19894759
PLoS One Circulating Levels of Fatty Acid-Binding Protein Family and Metabolic Phenotype in the General Population Pubmed: 24278421
PLOS ONE Transcriptome and Metabolome Analyses in Exogenous FABP4- and FABP5-Treated Adipose-Derived Stem Cells. pubmed:27936164
Sci Rep. Serum FABP5 concentration is a potential biomarker for residual risk of atherosclerosis in relation to cholesterol efflux from macrophages. pubmed:28303004
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