ELISA Kit for Peroxisome Proliferator Activated Receptor Gamma (PPARg)

PPAR-G; PPARG1; PPARG2; NR1C3; Glitazone Receptor; Nuclear Receptor Subfamily 1 Group C Member 3

Specificity

This assay has high sensitivity and excellent specificity for detection of Peroxisome Proliferator Activated Receptor Gamma (PPARg).
No significant cross-reactivity or interference between Peroxisome Proliferator Activated Receptor Gamma (PPARg) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Peroxisome Proliferator Activated Receptor Gamma (PPARg) and the recovery rates were calculated by comparing the measured value to the expected amount of Peroxisome Proliferator Activated Receptor Gamma (PPARg) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 91-98 94
heparin plasma(n=5) 82-98 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Peroxisome Proliferator Activated Receptor Gamma (PPARg) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Peroxisome Proliferator Activated Receptor Gamma (PPARg) 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 Peroxisome Proliferator Activated Receptor Gamma (PPARg) 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) 81-101% 94-105% 90-98% 98-105%
EDTA plasma(n=5) 92-101% 94-101% 79-93% 94-101%
heparin plasma(n=5) 97-105% 97-105% 79-90% 83-93%

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
Neuroscience Letters Elevated levels of PPAR-gamma in the cerebrospinal fluid of patients with multiple sclerosis Pubmed: 24021801
J Cell Biochem. Nonivamide enhances miRNA let‐7d expression and decreases adipogenesis PPARγ expression in 3T3‐L1 cells Pubmed:25704235
Osteoarthritis Cartilage Establishment of a rabbit model to study the influence of advanced glycation end products accumulation on osteoarthritis and the protective effect of pioglitazone PubMed: 26321377
J Clin Diagn Res. Evaluation of Protein Kinase Cβ and PPARγ Activity in Diabetic Rats Supplemented with Momordica charantia pmc:PMC4866090
Journal of Clinical&Diagnostic Research Evaluation of Protein Kinase Cβ and PPARγ Activity in Diabetic RatsSupplemented with Momordica charantia. pubmed:27190792
Osteoarthritis and Cartilage Establishment of a rabbit model to study the influence of advanced glycation end productsaccumulation on osteoarthritis and the protective effect of pioglitazone. pubmed:26321377
Neuroscience Letters Unlike PPARgamma, neither other PPARs nor PGC-1alpha is elevated in the cerebrospinal fluid of patients with multiple sclerosis pubmed:28483651
European Journal of Immunology Engulfment of Hb‐activated platelets differentiates monocytes into pro‐inflammatory macrophages in PNH patients Pubmed:29677388
Histochemistry and Cell Biology Simpson–Golabi–Behmel syndrome human adipocytes reveal a changing phenotype throughout differentiation Pubmed:29574488
molecular and cellular biochemistry Maternal omega-3 fatty acids and vitamin E improve placental angiogenesis in late-onset but not early-onset preeclampsia Pubmed: 31420792
Nutrients.  Hyperglycemia Changes Expression of Key Adipogenesis Markers (C/EBPα and PPARᵞ) and Morphology of Differentiating Human Visceral Adipocytes Pubmed: 31398873
journal of biochemical and molecular toxicology Indomethacin and juglone inhibit inflammatory molecules to induce apoptosis in colon cancer cells Pubmed: 31916655
J Cosmet Dermatol In©\vitro effect of pine bark extract on melanin synthesis, tyrosinase activity, production of endothelin©\1 and PPAR in cultured melanocytes exposed to Ultraviolet?¡­ 33960120
American Chemical Society TRPA1 Agonist Cinnamaldehyde Decreases Adipogenesis in 3T3-L1 Cells More Potently than the Non-agonist Structural Analog Cinnamyl Isobutyrate 33403292
PLoS One Quantitative real-time measurement of endothelin-1-induced contraction in single non-activated hepatic stellate cells 34343209
Prostaglandins Leukot Essent Fatty Acids Maternal Vitamin D Deficiency Reduces Docosahexaenoic Acid, Placental Growth Factor and Peroxisome Proliferator Activated Receptor Gamma levels in the Pup … 34768025
Food and Chemical Toxicology Melatonin attenuates cisplatin-induced acute kidney injury in mice: Involvement of PPARα and fatty acid oxidation Pubmed:35367536
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