High Sensitive ELISA Kit for Apolipoprotein M (APOM)

Apo-M; G3a; NG20; Protein G3a

Specificity

This assay has high sensitivity and excellent specificity for detection of High Sensitive Apolipoprotein M (APOM).
No significant cross-reactivity or interference between High Sensitive Apolipoprotein M (APOM) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant High Sensitive Apolipoprotein M (APOM) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Apolipoprotein M (APOM) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-95 89
EDTA plasma(n=5) 98-105 101
heparin plasma(n=5) 80-88 83

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Apolipoprotein M (APOM) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Apolipoprotein M (APOM) 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 High Sensitive Apolipoprotein M (APOM) 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) 94-104% 88-101% 89-103% 79-94%
EDTA plasma(n=5) 94-105% 85-102% 97-105% 89-96%
heparin plasma(n=5) 92-105% 79-102% 83-97% 82-101%

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
Joint Bone Spine Apolipoprotein m (APOM) levels and APOM rs805297 G/T polymorphism are associated with increased risk ofrheumatoid arthritis Pubmed: 23660425
Apoptosis The miR-573/apoM/Bcl2A1-dependent signal transduction pathway is essential for hepatocyte apoptosis and hepatocarcinogenesis PubMed: 26201458
Lipids in Health and Disease Effects of hyperlipidaemia on plasma apolipoprotein M levels in patients with type 2 diabetes mellitus: an independent case–control study pubmed:27633510
Lipids in Health and Disease A single-nucleotide polymorphism C-724 /del in the proter region of the apolipoprotein M gene is associated with type 2 diabetes mellitus pubmed:27576735
clinica chimica acta ApoM/HDL-C and apoM/apoA-I ratios are indicators of diabetic nephropathy in healthy controls and type 2 diabetes mellitus. pubmed:28073663
International Journal of Clinical and Experimental Pathology Level of serum apolipoprotein m and its diagnostic significance in inflammatory bowel disease files:ijcep0033416.pdf
Scandinavian Journal of Medicine & Science in Sports Endurance training selectively increases HDL-bound sphingosine-1-phosphate in the plasma pubmed:28493600
INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY HDL-S1P Protects Endothelial Function and Reduces Lung Injury During Sepsis in vivo and in vitro Pubmed: 32750426
Biology-Basel Biomechanical Properties of Blood Plasma Extracellular Vesicles Revealed by Atomic Force Microscopy 33374530
researchsquare Apolipoprotein M in High Density Lipoprotein Protects Against Astrocyte Apoptosis Induced By Ischemic Insult Via Sphingosine 1-Phosphate Signaling
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