ELISA Kit for Anti-Pentraxin 3, Long Antibody (Anti-PTX3)

TNFAIP5; TSG-14; Pentaxin-Related Gene,Rapidly Induced By IL-1 Beta; Tumor Necrosis Factor Alpha-Induced Protein 5; Tumor necrosis factor-inducible gene 14 protein

Specificity

This assay has high sensitivity and excellent specificity for detection of Anti-Pentraxin 3, Long Antibody (Anti-PTX3).
No significant cross-reactivity or interference between Anti-Pentraxin 3, Long Antibody (Anti-PTX3) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Anti-Pentraxin 3, Long Antibody (Anti-PTX3) and the recovery rates were calculated by comparing the measured value to the expected amount of Anti-Pentraxin 3, Long Antibody (Anti-PTX3) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 91-102 97
EDTA plasma(n=5) 84-97 90
heparin plasma(n=5) 95-102 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Anti-Pentraxin 3, Long Antibody (Anti-PTX3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Anti-Pentraxin 3, Long Antibody (Anti-PTX3) 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 Anti-Pentraxin 3, Long Antibody (Anti-PTX3) 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) 80-98% 91-98% 98-105% 95-104%
EDTA plasma(n=5) 92-104% 78-90% 81-94% 81-88%
heparin plasma(n=5) 81-94% 87-95% 81-97% 97-105%

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
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 5 times;
5. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
6. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Mediators of Inflammation Biochemical Analysis of Pentraxin 3 and Fibrinogen Levels in Experimental Periodontitis Model PubMed: 22966213
Ibnosina Journal of Medicine and Biomedical Sciences Effect of Vagus Nerve Stimulation on Focal Transient Cerebral Ischemia and Reperfusion in Adult Male White New Zealand Rabbits Sfu: Source
Hemodialysis International Hemodialysis-induced changes in the blood composition affect function of the endothelium Onlinelibrary: hdi.12148
Hemodial Int. Hemodialysis‐induced changes in the blood composition affect function of the endothelium Pubmed:24612443
Journal of Obstetrics and Gynaecology Research The effects of sunitinib on endometriosis Pubmed:25093747
Gynecol Endocrinol. Exenatide improves ovarian and endometrial injury and preserves ovarian reserve in streptozocin induced diabetic rats Pubmed:25366063
Taiwanese Journal of Obstetrics and Gynecology Ovarian failure in diabetic rat model: Nuclear factor-kappaB, oxidative stress, and pentraxin-3 Pubmed:25510691
Journal of Obstetrics and Gynaecology Research Octreotide protects ovary against ischemia–reperfusion injury in rats: Evaluation of histological and biochemical parameters PubMed: 26223286
J Obstet Gynaecol The long pentraxin-3 is a useful marker for diagnosis of ovarian torsion: An experimental rat model PubMed: 26470829
Neurology Research International Accumulation of α-Synuclein in Cerebellar Purkinje Cells of Diabetic Rats and Its Potential Relationship with Inflammation and Oxidative Stress Markers 5952149
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