ELISA Kit for Lipoxin A4 (LXA4)

LX-A4

Specificity

This assay has high sensitivity and excellent specificity for detection of Lipoxin A4 (LXA4).
No significant cross-reactivity or interference between Lipoxin A4 (LXA4) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Lipoxin A4 (LXA4) and the recovery rates were calculated by comparing the measured value to the expected amount of Lipoxin A4 (LXA4) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 95-103 99
EDTA plasma(n=5) 80-96 86
heparin plasma(n=5) 87-94 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Lipoxin A4 (LXA4) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Lipoxin A4 (LXA4) 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 Lipoxin A4 (LXA4) 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) 80-101% 96-104% 81-99% 86-103%
EDTA plasma(n=5) 81-91% 80-88% 98-105% 89-97%
heparin plasma(n=5) 79-88% 93-101% 84-92% 87-94%

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 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

Assay procedure summary

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

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Magazine Citations
Neurotoxicology Redox modulation of cellular stress response and lipoxin A4 expression by Coriolus versicolor in rat brain: Relevance to Alzheimer&#039;s disease pathogenesis PubMed: 26433056
Immunity & Ageing Redox modulation of cellular stress response and lipoxin A4 expression by Hericium Erinaceus in rat brain: relevance to Alzheimer's disease pathogenesis. pubmed:27398086
BMC cardiovascular disorders Acute coronary syndrome and acute kidney injury: role of inflammation in worsening renal function pubmed:28747177
Liver transplantation  CDP‐choline protects liver from ischemia/reperfusion injury preserving mitochondrial function and reducing oxidative stress Pubmed:29679463
Journal of Immunology Leukocyte CD300a Contributes to the Resolution of Murine Allergic Inflammation Pubmed: 30315138
Experimental and Therapeutic Medicine Dynamic changes and clinical significance of LXA4 in the perioperative period of cardiopulmonary bypass
Biochem Genet Lipoxin A4-Mediated p38 MAPK Signaling Pathway Protects Mice Against Collagen-Induced Arthritis 33221976
The mimetic peptide Ac2-26 of annexin A1 attenuates inflammation and apoptosis in sepsis-induced acute kidney injury
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