- Regulatory Status
- Other Names
- TRAP, TRACP, TRACP5a, TRACP5b, Tartrate-Resistant Acid ATPase, Acid Phosphatase 5 Tartrate Resistant, Type 5 Acid Phosphatase
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- Product Citations
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Tartrate-resistant acid phosphatase (TRAP/ACP5) is a metalloenzyme, categorized as an acid phosphatase, and catalyzes the transformation of orthophosphoric monoester to alcohol and orthophosphate. It is an iron-containing glycoprotein that is synthesized as a monomeric proenzyme of 35 kD. ACP5 plays a role in bone resorption and is secreted by active osteoclasts. It dephosphorylates osteopontin (a bone matrix protein) and subsequently, promotes osteoclast detachment and migration. ACP5 expression correlates with tumor progression in many types of cancer such as colorectal cancer, ovarian cancer, breast cancer, and melanoma. Expression of ACP5, in colorectal cell lines in vitro, promotes tumor cell proliferation and invasion. ACP5 in serum has been used as a marker for broad bone metastasis. In addition, ACP5 removes the mannose 6-phosphate which is the recognition marker from lysosomal proteins.Product Details
- Mouse ACP5, amino acid Thr22-Pro327 (Accession # Q05117) was expressed in CHO cells. The carboxy terminus contains an GGS-6His tag.
- Molecular Mass
- The 315 amino acid recombinant protein has a predicted molecular mass of approximately 35.4 kD. The DTT-reduced and non-reduced protein migrates at approximately 40 kD by SDS-PAGE. The predicted N-terminal amino acid is Thr.
- >95%, as determined by Coomassie stained SDS-PAGE.
- 0.22 µm filtered protein solution is in PBS, pH 7.2
- Endotoxin Level
- Less than 0.10 EU per µg protein as determined by the LAL method.
- 10 and 25 µg sizes are bottled at 200 µg/mL.
- Storage & Handling
- Unopened vial can be stored at -20°C or -70°C for six months. For maximum results, quick spin vial prior to opening. Avoid repeated freeze/thaw cycles.
- Mouse ACP5 has the ability to cleave the substrate, p-Nitrophenyl phosphate (pNPP). The specific activity is >22,000 pmol/min/µg.
- Application Notes
ALP5 Assay Procedure
- In assay buffer, prepare dilution of mouse ACP5 at 0.1 µg/mL
- Add 50 µL of mACP5 and 50 µL of prepared 2 mM substrate to plate. Include a blank for the substrate containing 50 µL assay buffer and 50 µL of 2 mM substrate.
- At room temperature, incubate plate for 10 minutes in the dark.
- To develop the color and stop the reaction, add 100 µL of 0.2 M NaOH.
- Read (top read) absorbance in endpoint mode at 410 nm.
- Calculate the specific activity:
Specific Activity (pmol/min/µg) = Adjusted Abs* (OD) x Conversion Factor** (pmol/OD)
Incubation time (min) x amount of enzyme (µg)
*Adjusted for the substrate blank
**Derived using calibration standard p-Nitrophenol (Sigma-Aldrich, Catalog # 241326).
- Recombinant mACP5: 0.005 µg
- Substrate: 0.5 mM
- Assay Buffer: 50 mM NaOAc, pH 5.0
- Mouse TRACP/PAP/ACP5
- Substrate: p-Nitrophenyl phosphate (Sigma-Aldrich, Catalog # N2765)
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- NaOH, 0.2 M in deionized water
BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at firstname.lastname@example.org.
Osteoclast, macrophages, dendritic cells, endometrial glandular epithe, cancer cells
- Normal bone development, osteoblast regulation, macrophage function, bone resorption, and osteoclast activity. RANKL can increase the expression of ACP5
- In vivo substrates: Osteopontin, Mannosa 6 Phosphate-Lysosomal proteins
- Mouse ACP5 has the ability to cleave the substrate, p-Nitrophenyl phosphate (pNPP).
- Cell Type
- Epithelial cells, Macrophages, Osteoclasts
- Biology Area
- Cancer Biomarkers, Cell Biology, Immuno-Oncology
- Molecular Family
- Enzymes and Regulators, Lysosomal Markers, Proteases
- Antigen References
- Suter A, et al. 2001. Development. 128:4899.
- Fretz JA, et al. 2008. Mol Endocrinol. 22:737.
- Sun P, et al. 2008. Proc Natl Acad Sci U S A. 105:16590.
- Ek-Rylander B & Andersson G. 2010. Exp Cell Res. 316:443.
- Reithmeier A, et al. 2017. BMC Cancer. 17:650.
- Bian ZQ, et al. 2019. Am J Cancer Res. 9:22.
- Gene ID
- 11433 View all products for this Gene ID
- View information about ACP5 on UniProt.org
- Why choose BioLegend recombinant proteins?
• Each lot of product is quality-tested for bioactivity as indicated on the data sheet.
• Greater than 95% Purity or higher, tested on every lot of product.
• 100% Satisfaction Guarantee for quality performance, stability, and consistency.
• Ready-to-use liquid format saves time and reduces challenges associated with reconstitution.
• Bulk and customization available. Contact us.
• Learn more about our Recombinant Proteins.
- How does the activity of your recombinant proteins compare to competitors?
We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!
- What is the specific activity or ED50 of my recombinant protein?
The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.
- Have your recombinants been tested for stability?
Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.
- Does specific activity of a recombinant protein vary between lots?
Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.
- How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
- Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL)