TRAP 5 Human Assay (Tartrate Resistant Acid Phosphatase 5, TRACP 5)

Research topic

Bone and cartilage metabolism

Features

• The total assay time is less than four hours.
• The kit measures only the TRAP 5 proteins exhibiting enzyme activityrecombinant protein based.
• No animal sera are used.

Storage/Shipping

Store the kit at 2–8°C. Under these conditions, the kit is stable until the expiration date (see label on the box).

Summary

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.

Assay format

Immunocapture enzyme activity assay

Sample requirements

25 µl/well

Applications

Plasma-EDTA, Serum

Calibration Curve

Calibration range

0.2 – 4 U/l

Limit of detection

Analytical Limit of Detection is calculated from the real TRAP 5 values in wells and is 0.4U/l

Limit of quantification

Assay Sensitivity takes the dilution of samples into consideration and is calculated according to the formula: Assay Sensitivity = Analytical Limit of Detection x sample dilution = 0.4U/l x 4 = 1.6 U/l

Intra-assay (Within-Run, n=8)

CV = 3.2%

Inter-assay (Run-to-Run, n=4)

CV = 6.2 %

Spiking Recovery

96%

Dilution Linearity

104 %

Cross-Reactivity

Human, Dog, Monkey, Rabbit

References to this product

• 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)

References to summary

• Minkin C. Bone acid phosphatase: tartrate-resistant acid phosphatase as a marker of osteoclast function. Calcif Tissue Int. 1982, 34, 285–290.
• Lau KH, Onishi T, Wergedal JE, Singer FR, Baylink DJ. Characterization and assay of tartrate-resistant acid phosphatase activity in serum: potential use to assess bone resorption. Clin Chem. 1987, 33, 458–462.
• Nakanishi M, Yoh K, Uchida K, Maruo S, Matsuoka A. Improved method for measuring tartrate-resistant acid phosphatase activity in serum. Clin Chem. 1998, 44, 221–225.
• Halleen JM, Alatalo SL, Suominen H, Cheng S, Janckila AJ, Vaananen HK. Tartrateresistant acid phosphatase 5b: a novel serum marker of bone resorption. J Bone Miner Res. 2000, 15, 133–1345.
• Lamp EC, Drexler HG. Biology of tartrate-resistant acid phosphatase. Leuk Lymphoma. 2000, 39, 477–484.
• Nakanishi M, Yoh K, Miura T, Ohasi T, Rai SK, Uchida K. Development of a kinetic assay for band 5b tartrate-resistant acid phosphatase activity in serum. Clin Chem. 2000, 46, 469–473.
• Waguespack SG, Hui SL, White KE, Buckwalter KA, Econs MJ. Measurement of tartrate-resistant acid phosphatase and the brain isoenzyme of creatine kinase accurately diagnose type II autosomal dominant osteopetrosis but does not identify gene carriers. J Clin Endocrinol Metab. 2002, 87, 2212–2217.
• Igarashi Y, Lee M, Matsuzaki S. Acid phosphatases as markers of bone metabolism. J Chromatogr B. 2002, 781, 345–358.
• Terpos E, de la Fuente J, Szydlo R, Hatjiharissi E, Viniou N, Meletis J, Yataganas X, Goldman JM, Rahemtulla A. Tartrate-resistant acid phosphatase isoform 5b: a novel serum markerfor monitoring bone disease in multiple myeloma. Int J Cancer. 2003, 106, 455–457.