Recombinant Human ACP6 Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human ACP6 Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a substrate, p-Nitrophenyl phosphate (pNPP). The specific activity is >1800 pmol/min/μg, as measured under the described conditions.
• Source: Chinese Hamster Ovary cell line, CHO-derived human ACP6 protein
  Glu33-Glu428, with a C-terminal 6-His tag
• Accession #: BAA91310
• N-terminal Sequence: Glu33
• Protein/Peptide Type: Recombinant Enzymes
• Gene: ACP6
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 46 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 43-45 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris and NaCl.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
• Assay Procedure:
  • Assay Buffer: 50 mM Sodium acetate, 0.05% Triton® X-100, pH 5.0
  • Recombinant Human ACP6 (rhACP6) (Catalog # 7766-AP)
  • Substrate: p-Nitrophenyl phosphate (Sigma, Catalog # N2765), 10 mM stock in deionized water
  • NaOH, 0.2 M in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhACP6 to 0.8 µg/mL in Assay buffer.
  2. Dilute Substrate to 2 mM in Assay buffer.
  3. In a plate, load 50 µL of 0.8 µg/mL rhACP6, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL of Assay buffer and 50 µL of 2 mM Substrate.
  4. Incubate plate at room temperature for 30 minutes in the dark.
  5. Add 100 µL of 0.2 M NaOH to all wells to stop the reaction and develop the color.
  6. Read plate at 410 nm (absorbance) in endpoint mode.
  7. Calculate 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 Substrate Blank
  **Derived using calibration standard p-Nitrophenol (Sigma, Catalog # 241326).

  Per Well:
  • rhACP6: 0.04 µg
  • pNPP: 0.5 mM

Notes

Coomassie is a registered trademark of Imperial Chemical Industries Ltd. Triton is a registered trademark of Union Carbide Corp.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human ACP6 Protein, CF

• acid phosphatase 6, lysophosphatidicPACPL1
• Acid phosphatase-like protein 1
• ACP6
• ACPL1
• ACPL1acid phosphatase like 1
• EC 3.1.3.2
• LPAP
• LPAPlysophosphatidic acid phosphatase type 6
• PACPL1

Background

Lysophosphatidic Acid Phosphatase Type 6 (ACP6) is a membrane-bound protein that belongs to the Histidine Acid Phosphatase Family. ACP6 is expressed in kidney, heart, small intestine, muscle, liver, prostate, testis, ovary, and colon tissues (1, 2). Its function is to hydrolyze lysophosphatidic acid (LPA) to form monoacylglycerol. LPA is one of the simplest of all known phospholipids which has a variety of biological functions including cell proliferation, migration, and survival (3). LPA is found at high levels in serum but also present in saliva, follicular fluid, seminal plasma and malignant effusions and is implicated in many cancers including ovarian, breast and prostate (3-5).

1. Hiroyama, M. and T. Takenawa (1999) J. Biol. Chem. 274:29172.
2. Takayama I. et al. (2002) Gut 50:790.
3. Mills, G. and W. Moolenaar (2003) Nature 3:582.
4. Xu, Y. et al. (1995) Biochem J. 309:933.
5. Xie, Y. et al. (2002) J. Biol. Chem. 277:32516.

Bioinformatics

• Gene Symbol: ACP6
• Uniprot:
  • BAA91310()