Released on = December 6, 2006, 9:56 am

Press Release Author = Albert Hong

Industry = Biotech

Press Release Summary = Protein phosphatases play key cellular regulatory roles in
such processes as differentiation, proliferation and apoptosis(1).


Press Release Body = As a result, they have received significant attention as
potential drug-screening targets. AnaSpec's range of protein phosphatase assay kits
include both fluorimetric and colorimetric readouts. The colorimetric assay kit
utilizes p-nitrophenyl phosphate (pNPP) as a substrate. It shows high sensitivity
and a wide linear range. The detection limit is generally 3 ng or lower. The
fluorimetric assays are even more sensitive than the colorimetic assay, and there
are two kits to choose from. The EnzoLyteTFDP Assay Kit, using FDP (3,6-Fluorescein
Diphosphate), is optimized for assaying protein phosphatases which require pH>6.5
for optimal activity, while the EnzoLyteT MFP Assay Kit is for phosphatases which
require pH< 6.5. Both fluorophores emit at about 520 nm upon dephosphorylation.

EnzoLyteT FDP Protein Phosphatase Assay Kit
Type: Fluorimetric, Abs (nm) 485±20, Em (nm) 528±20, Cat# 71100

EnzoLyteT MFP Protein Phosphatase Assay Kit
Type: Fluorimetric, Abs (nm) 488±20, Em (nm) 520±20, Cat# 71104

EnzoLyteT pNPP Protein Phosphatase Assay Kit
Type: Colorimetric, Abs (nm)405, Em (nm) NA, Cat# 71105

The change in alkaline phosphatase level and activity is involved in a variety of
physiological and pathological events, such as bone development, (2 )bone-related
diseases(3), gestation related diseases(4), inflammatory bowel disease(5),
post-parathyroidectomy stage(6), and drug toxicity(7). Alkaline phosphatase is also
widely used, when conjugated to a secondary antibody, as a reporter in ELISA assays.
The Enzolyte Alkaline Phosphatase Assay Kits are designed to detect enzyme activity
in biological samples, while the Enzolyte Alkaline Phosphatase ELISA Assay Kits are
optimized to detect alkaline phosphatase in ELISA.

EnzoLyteT FDP Alkaline Phosphatase Assay Kit
Type: Fluorimetric, Abs (nm) 485±20, Em (nm) 528±20, Cat # 71109

EnzoLyteT pNPP Alkaline Phosphatase Assay Kit
Type: Colorimetric, Abs (nm) 405, Em (nm) NA, Cat # 71230

EnzoLyteT FDP Alkaline Phosphatase ELISA Assay Kit (AP-conjugated Goat anti-rabbit
IgG included in kit)
Type: Fluorimetric, Abs (nm) 485±20, Em (nm) 528±20, Cat # 71101-R

EnzoLyteT FDP Alkaline Phosphatase ELISA Assay Kit (AP-conjugated Goat anti-mouse
IgG included in kit)
Type: Fluorimetric, Abs (nm) 485±20, Em (nm) 528±20, Cat # 71101-M

EnzoLyteT pNPP Alkaline Phosphatase Assay Kit
Type: Colorimetric, Abs (nm) 405, Em (nm) NA, Cat # 71230

The placental alkaline phosphatase is the most stable isoenzyme among the four
mammalian alkaline phosphatase isoenzymes, and it only exists naturally in the
placenta of higher primates. These characteristics make placental alkaline
phosphatase the alkaline phosphatase of choice to serve as a reporter gene for the
analysis of promoter activity and gene expression in cell culture or animals. The
natural form of placental alkaline phosphates is membrane-anchored. The recombinant
form of placental alkaline phosphatase, secreted alkaline phosphatase (SEAP), can be
efficiently secreted into tissue culture medium and serum.8,9 This unique
characteristic of SEAP provides the capability of performing continuous kinetic
analysis of gene expression over a period of time using a single cell culture or
animal.

The EnzoLyteT FDP Secreted Alkaline Phosphatase Reporter Gene Assay Kits assay both
secreted and membrane-bound placental alkaline phosphatase. The fluorimetric assay
kit uses FDP as the fluorogenic phosphatase substrate. Fluorescein, the final
hydrolytic product of FDP, has a very high extinction coefficient and emission
quantum yield. As a result, the assay is more sensitive than the colorimetric kit,
which uses pNPP. Using FDP can detect as low as 0.1 ng of phosphatase with a linear
range of three orders of magnitude, while the limit of detection using pNPP is 10
ng.

EnzoLyteT FDP SEAP Assay Kit
Type: Fluorimetric, Abs (nm) 485±20, Em (nm) 528±20, Cat # 71108

EnzoLyteT pNPP SEAP Assay Kit
Type: Colorimetric, Abs (nm) 405, Em (nm) NA, Cat # 71233

The change in acid phosphatase level and activity is involved in a variety of
physiological and pathological events, such as prostate puberty, rheumatoid
arthritis (10) bone-resorption related diseases (11) and diabetes (12). Acid
phosphatase is also a serum marker of tumor bone metastasis (13, 14).
The EnzoLyteT MFP Acid Phosphatase Assay Kit is optimized to measure acid
phosphatase activities using MFP as a fluorogenic substrate.

EnzoLyteT MFP Acid Phosphatase Assay Kit
Type: Fluorimetric, Abs (nm) 488±20, Em (nm) 520±20, Cat # 71231

The EnzoLyteT MG Phosphate Assay Kit is based on the quantification of the
blue-green complex formed between malachite green (MG) molybdate and free phosphate.
This non-radioactive colorimetric assay kit has been optimized to offer superior
sensitivity with a detection limit of as low as 8 pmoles of phosphate and an
extended linear range of 0.1-50 μM phosphate. The assay has been widely used to
quantify liberated phosphate in phosphatase assays, lipase assays, and nucleotide
triphosphatase assays.

EnzoLyteT MG Phosphate Assay Kit
Type: Colorimetric, Abs (nm) 405, Em (nm) NA, Cat # 71103

For more information visit our website.

References:
1. Kupcho, K. et al. J. Biomol. Screening 9, 223 (2004).
2. Kotobuki, N. et al. Cell Transplant. 13, 377 (2004).
3. Wyckoff, MH. et al., J. Clin. Endocrinol. Metab. 90, 1233 (2005).
4. Boronkai, A. et al., J.Clin.Pathol. 58, 72-76 (2005).
5. Sanchez, DM. et al., Biochem. Pharmacol. 68, 2317 (2004).
6. Morrone, LF. et al. Ann. Ital. Med. Int. 19, 189 (2004).
7. Papaldo, P. et al., Cancer Invest 22, 650 (2004).
8. Berger, J. et al. Gene 66, 1 (1988).
9. Cullen, BR. and MH. Malim, Methods Enzymol. 216, 362 (1992).
10. Hayman, AR. and TM. Cox, Cell Biochem. Funct. 22, 275 (2004).
11. Janckila, AJ. et al. J. Leukoc. Biol. 77, 209 (2005).
12. Bottini, N. et al. Metabolism 53, 995 (2004).
13. Chao, TY. et al., J. Biomed. Sci. 11, 511 (2004).
14. Seki, K. et al., Am. J. Surg. Pathol. 28, 1384 (2004).


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