Recombinant Human ACP6 Protein, CF

Catalog #: 7766-AP Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
7766-AP-010

R&D Systems Recombinant Proteins and Enzymes

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Recombinant Human ACP6 Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

Activity

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
Analysis

Glu33

Predicted Molecular Mass

46 kDa

SDS-PAGE

43-45 kDa, reducing conditions

Product Datasheets

7766-AP

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

7766-AP

Formulation	Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Assay Procedure

Materials

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

Dilute rhACP6 to 0.8 µg/mL in Assay buffer.
Dilute Substrate to 2 mM in Assay buffer.
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.
Incubate plate at room temperature for 30 minutes in the dark.
Add 100 µL of 0.2 M NaOH to all wells to stop the reaction and develop the color.
Read plate at 410 nm (absorbance) in endpoint mode.
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

Reconstitution Calculator

Background: ACP6

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).

References

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

Long Name

Lysophosphatidic Acid Phosphatase 6

Entrez Gene IDs

51205 (Human); 66659 (Mouse); 295305 (Rat)

Alternate Names

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