TRAP 5 Human Assay

Regulatery Status: RUO
Type: Immunocapture enzyme activity assay
Other Names Status: TRACP 5, Tartrate Resistant Acid Phosphatase 5
Species: Human
Catalog No Size
Product Catalog No: RD197025000 Pack Size: 96 wells (1 kit)

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Product Features

TRAP 5 (serum band 5 tartrate-resistant acid phosphatase, TRACP 5; EC 3.1.3.2) is a glycoprotein of 35–37 kDa. TRAP 5 belongs to the most abundant enzymes in osteoclasts. It is expressed in certain differentiated cells of the mononuclear phagocyte system, particularly osteoclasts and alveolar macrophages, where it takes an active part in bone resorption process. High blood levels of TRAP 5 are usually associated with active bone remodelling. Increased serum levels are observed during normal bone growth among healthy children. Elevated serum TRAP levels have been detected in diseases characterized by increased bone resorption; Paget’s disease of the bone, hemodialysis, primary hyperparathyro­idism, metastatic malignancies involving bone resorption, multiple myeloma and bilaterally ovariectomized women. Post-menopausal women have higher levels of serum than post-menopausal women on estrogen replacement therapy. Therefore specific determination of TRAP 5 activity can be essential for clinical assessment of bone metabolism.

Technical Sheet / Info

Intended use

The RD197025000 Human TRAP 5 Assay is an ImmunoCapture Enzyme-Activity Assay for the quantitative measurement of human TRAP 5 in serum and EDTA plasma.

  • The total assay time is less than 4 hours
  • The kit measures TRAP 5 proteins exhibiting enzyme activity
  • Assay format is 96 wells
  • Quality Controls are recombinant TRAP 5 protein based. No animal sera are used
  • Calibrators are recombinant protein based
  • Components of the kit are provided ready to use, concentrated or lyophilized

Clinical Application

  • Bone and cartilage metabolism

Test principle

In the BioVendor Human TRAP 5 Assay, Calibrators, Quality Control and samples are incubated in microplate wells pre-coated with monoclonal anti-human TRAP 5 antibody. After a thorough wash, TRAP 5 bound to the antibody is allowed to react with the pNPP substrate at pH 5.5. The reaction is stopped by addition of hydroxide solution and absorbance of the resulting yellow colour product is measured spectrophotometrically at 405 nm. The absorbance is proportional to the enzymatic activity of TRAP 5. A standard curve is constructed by plotting absorbance values versus enzyme activities of recombinant TRAP 5 calibrators, and enzyme activity of unknown samples are determined (U/l) using this standard curve.

Summary of protocol

  • Reconstitute Calibrators
  • Reconstitute QC, dilute Samples and QC 4×
  • Add 100 μl Calibrators, QC and samples
  • Add 50 μl Dissociating Solution
  • Incubate at RT for 2 hours/300 rpm
  • Wash plate 3 times
  • Add 100 μl Substrate Solution
  • Incubate at 37°C for 1.5 hour/no shaking!
  • Add 100 μl Stop Solution
  • Read absorbance and calculate results

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    References

    – Halleen JM, Tiitinen SL, Ylipahkala H, Fagerlund KM, Vaananen HK. Tartrate-resistant Acid Phosphatase 5b (TRACP 5b) as a Marker of Bone Resorption. Clin Lab . 52(9-10):499-509 (2006)

    – Monier-Faugere MC, Mawad H, Malluche HH. Opposite effects of calcitriol and paricalcitol on the parathyroid hormone-(1-84)/large carboxy-terminal-parathyroid hormone fragments ratio in patients with stage 5 chronic kidney disease. Clin J Am Soc Nephrol. 2007 Nov;2 (6):1255-60

    – Taylan A, Sari I, Akinci B, Bilge S, Kozaci D, Akar S, Colak A, Yalcin H, Gunay N, Akkoc N. Biomarkers and cytokines of bone turnover: extensive evaluation in a cohort of patients with ankylosing spondylitis. BMC Musculoskelet Disord. 2012;13:191

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