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Signal Guard™ phosphatase reaction stopping solution

Alkaline and acid phosphatases are the hydrolase enzymes responsible for dephosphorylation of molecules such as nucleotides, proteins and alkaloids under alkaline and acidic conditions, respectively. Some phosphatases assays and tests require the phosphatase reactions to be stopped for further analysis using the stop solutions. Our Signal Guard™ phosphatase reaction stopping solution is a ready to use reagent that provides a convenient tool for terminating fluorescence and colorimetric signal-generating phosphatase reactions at a user-determined time point, and also keep the fluorescence or colorimetric signal stable for up to 18 hours. The Signal Guard™ phosphatase reaction stopping solution is optimized and compatible with alkaline, acid and protein phosphatase. Compared to other commercial phosphatase stopping reagents, our Signal Guard™ phosphatases reaction stopping solution is nearly neutral and mild. It is compatible with a vast majority of colorimetric and fluorimetric phosphatase substrates while the stop reagents from other vendors are not compatible with most of fluorescence-based phosphatase assays due to their extremely high pH.

Example protocol

SAMPLE EXPERIMENTAL PROTOCOL

Assay Protocol for one 96-well plate

  1. Warm the Signal Guard™ phosphatase reaction stopping solution to room temperature.

  2. At the desired stopping time point, add 50 µL of Signal Guard™ phosphatase reaction stopping solution per 200 µL volume in each microplate well. Note: For other reaction volumes, adjust the addition of Signal Guard™ Phosphatase reaction stopping solution proportionally (e.g. add 5 µL to a 25 µL reaction volume). The signal level should remain stable for at least 18 hours.

Citations

View all 104 citations: Citation Explorer
Discoveries of the phosphatidate phosphatase genes in yeast published in the Journal of Biological Chemistry
Authors: Carman, G. M.
Journal: J Biol Chem (2019): 1681-1689
The multiple functions of protein phosphatase 6
Authors: Ohama, T.
Journal: Biochim Biophys Acta Mol Cell Res (2019): 74-82
Tuning the Protein Phosphorylation by Receptor Type Protein Tyrosine Phosphatase Epsilon (PTPRE) in Normal and Cancer Cells
Authors: Liang, J., Shi, J., Wang, N., Zhao, H., Sun, J.
Journal: J Cancer (2019): 105-111
Dynamic structural rearrangements and functional regulation of voltage-sensing phosphatase
Authors: Sakata, S., Okamura, Y.
Journal: J Physiol (2019): 29-40
Redox inhibition of protein phosphatase PP2A: Potential implications in oncogenesis and its progression
Authors: Raman, D., Pervaiz, S.
Journal: Redox Biol (2019): 101105

References

View all 33 references: Citation Explorer
Enzyme-nanoparticle functionalization of three-dimensional protein scaffolds
Authors: Hill RT, Shear JB.
Journal: Anal Chem (2006): 7022
TNP-8N3-ADP photoaffinity labeling of two Na,K-ATPase sequences under separate Na+ plus K+ control
Authors: Ward DG, Taylor M, Lilley KS, Cavieres JD.
Journal: Biochemistry (2006): 3460
Xenopus apyrase (xapy), a secreted nucleotidase that is expressed during early development
Authors: Devader C, Webb RJ, Thomas GM, Dale L.
Journal: Gene (2006): 135
Immunoassay for B. globigii spores as a model for detecting B. anthracis spores in finished water
Authors: Farrell S, Halsall HB, Heineman WR.
Journal: Analyst (2005): 489
Controlled initiation of enzymatic reactions in micrometer-sized biomimetic compartments
Authors: Karlsson A, Sott K, Markstrom M, Davidson M, Konkoli Z, Orwar O.
Journal: J Phys Chem B Condens Matter Mater Surf Interfaces Biophys (2005): 1609
Page updated on December 17, 2024

Ordering information

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Unit size
Catalog Number11622
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Additional ordering information

Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
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Purchase orderSend to sales@aatbio.com
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Request quotation

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Refrigerated (2-8 °C)
UNSPSC12171501
The application of Signal Guard™ phosphatase reaction stopping solution in Acid phosphatase fluorescence (AAT Bioquest Cat#11627) assay. Two parallel reactions containing 100 mU/mL Acid phosphatase was initiated by adding 200 μL reaction mixture. Reactions were incubated at room temperature for times indicated above and then 50 µL Signal Guard™ phosphatase reaction stopping solution was added to one reaction. The plots demonstrated that the reaction is completely inhibited by Signal Guard™ phosphatase reaction stopping solution.
The application of Signal Guard™ phosphatase reaction stopping solution in Acid phosphatase fluorescence (AAT Bioquest Cat#11627) assay. Two parallel reactions containing 100 mU/mL Acid phosphatase was initiated by adding 200 μL reaction mixture. Reactions were incubated at room temperature for times indicated above and then 50 µL Signal Guard™ phosphatase reaction stopping solution was added to one reaction. The plots demonstrated that the reaction is completely inhibited by Signal Guard™ phosphatase reaction stopping solution.
The application of Signal Guard™ phosphatase reaction stopping solution in Acid phosphatase fluorescence (AAT Bioquest Cat#11627) assay. Two parallel reactions containing 100 mU/mL Acid phosphatase was initiated by adding 200 μL reaction mixture. Reactions were incubated at room temperature for times indicated above and then 50 µL Signal Guard™ phosphatase reaction stopping solution was added to one reaction. The plots demonstrated that the reaction is completely inhibited by Signal Guard™ phosphatase reaction stopping solution.
The application of Signal Guard™ phosphatase reaction stopping solution in Alkaline phosphatase colorimetric assay (AAT Bioquest Cat#11950). Two parallel reactions containing 100 mU/mL Alkaline phosphatase was initiated by adding 200 μL reaction mixture. Reactions were incubated at room temperature for times indicated above and then 50 µL Signal Guard™ phosphatase reaction stopping solution was added to one reaction. The plots demonstrated that the reaction is completely inhibited by Signal Guard™ phosphatase reaction stopping solution.

Documents

pdfSafety data sheet
pdfProduct protocol
pdfCertificate of analysis
Alkaline Phosphatase (ALP)
Signal Guard™ HRP conjugate stabilizer
Signal Guard™ HRP reaction stopping reagent