ELISA Kit for Thymidine Phosphorylase (TP)

TYMP; PDECGF; ECGF1; MNGIE; TP; HPD-ECGF; TdRPase; Platelet-Derived Endothelial Cell Growth Factor; Gliostatin

Specificity

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

Recovery

Matrices listed below were spiked with certain level of recombinant Thymidine Phosphorylase (TP) and the recovery rates were calculated by comparing the measured value to the expected amount of Thymidine Phosphorylase (TP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-94 85
EDTA plasma(n=5) 88-102 98
heparin plasma(n=5) 85-99 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Thymidine Phosphorylase (TP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Thymidine Phosphorylase (TP) 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 Thymidine Phosphorylase (TP) 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) 89-96% 98-105% 89-103% 88-96%
EDTA plasma(n=5) 94-105% 96-105% 78-89% 87-101%
heparin plasma(n=5) 78-96% 82-96% 80-101% 83-103%

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
Eur J Obstet Gynecol Reprod Biol. Profiling of selected angiogenesis-related genes in proliferative eutopic endometrium of women with endometriosis Pubmed: 24188612
PLoS One. Liver as a source for thymidine phosphorylase replacement in mitochondrial neurogastrointestinal encephalomyopathy Pubmed:24802030
BMC Clinical Pathology Thymidine phosphorylase expression is associated with time to progression in patients with metastatic colorectal cancer Pubmed:24936150
Oncotarget Capecitabine reverses tumor escape from anti-VEGF through the eliminating CD11bhigh/Gr1high myeloid cells Pubmed:29707135
Clinica Chimica Acta Prognostic significance of serum translocator protein in patients with traumatic brain injury Pubmed: 30385279
JOURNAL OF INTERNATIONAL MEDICAL RESEARCH Usefulness of postoperative serum translocator protein as a predictive marker for delirium after breast cancer surgery in elderly women Pubmed: 32529881
Bioengineering & Translational Medicine Targeting thymidine phosphorylase as a potential therapy for bone loss associated periprosthetic osteolysis
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