太鼎生物科技/太鼎食安科技

【如何建立快速藥物篩選系統】

PiColorLock採用超高靈敏度Malachite green磷酸根呈色方法，克服傳統呈色方法,低靈敏度限制。測量目標樣本中，是否含有ATPases、phosphatases、GTPases、nucleotidases酵素活性。

Innova Biosciences PiColorLock 針對實驗初篩所設計，可探討酵素活性是否扮演transport、signal transduction, protein biosynthesis及cell differentiation之角色。應用於ATPase、GTPase、Proteinases活性分析。提供high-throughput screen 大量ATPase或GTPase inhibitor。

PiColorLock產品，有分為Gold (靈敏度4~40uM)及ALS(靈敏度20~200uM)。

inorganic phosphate detection

產品特點

1.      PiColorLock分析套組，包括stabilizing solution，可避開acid labile substrate所造成的高背景值，其背景值很低( 1mM )。

2.      其分析波段為600~660nm，避開一般colored compounds之干擾。

3.      Pi-dye複合物極為穩定，其訊號強度可維持數小時之久。

4.      Superior呈色系統。採用Malachite Green磷酸根呈色方式，呈色靈敏度比傳統呈色方法高，克服低靈敏度的限制。

5.      適用於大部份的assay buffer。

6.      蛋白質濃度高時，不會抑制顏色的形成。

High Throughput Colorimetric ATPase Assays

This non radioactive colorimetric assay kit uses a 96 well format, and all the necessary reagents are supplied for measuring enzyme activity and are ideal for high throughput drug screening.

High Throughput Colorimetric GTPase Assays

This non radioactive colorimetric assay kit uses a 96 well format, and all the necessary reagents are supplied for measuring enzyme activity and are ideal for high throughput drug screening.

Pi Colorlock (ALS)

PiColorLock is a phosphate detection reagent for measuring Phosphatases, ATPases and Nucleotidases.

Pi Colorlock (Gold)

PiColorLock is a phosphate detection reagent for measuring Phosphatases, ATPases and Nucleotidases.

Colorimetric assays for ATPases are invariably based on the formation of colored complexes between an inorganic phosphate and a dye molecule under acidic conditions. Such assays are beset with problems of reagent precipitation and high backgrounds caused by impure substrates (i.e. contaminated with inorganic phosphate) and/or by non-enzymatic (acid) hydrolysis of the substrate.

At the heart of our assay kits is PiColorLock Gold, a superior phosphate detection reagent which ensures high stability of the colored dye-phosphate complexes. Together with specially purified substrates and proprietary stabilizers our kits offer the lowest possible assay backgrounds and outstanding assay performance.

Uniquely, Innova Biosciences ATPase assay kits contain highly purified ATP, which have had any free inorganic phosphate removed – this means the assays have the lowest possible background signal.

PiColorLock reagent may also be purchased separately for developing simple assays for any phosphate-generating enzyme. It is offered in both high (Gold) and standard sensitivity (ALS) formulations to suit almost any assay situation.

更多參考內容:

原廠連結: http://www.innovabiosciences.com/

文獻發表

ATPase Assay Kit

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Product

• Caveolin-1 regulates proliferation and
  osteogenic differentiation of human mesenchymal stem cells
  - Journal of Cellular
  Biochemistry (2012)


• Common Pathogenic Effects of Missense Mutations
  in the P-Type ATPase ATP13A2 (PARK9) Associated with Early-Onset
  Parkinsonism -
  PLOS One (2012)


• Structure of the VirB4 ATPase, alone and bound
  to the core complex of a type IV secretion system - PNAS (2012)


• Peroxisomal proteostasis involves a Lon family
  protein that functions as protease and chaperone - JBC (2012)


• Xeroderma pigmentosum group F protein binds to
  Eg5 and is required for proper mitosis: implications for XP-F and XFE
  - Genes to Cells (2012)


• hSWS1·SWSAP1 Is an
  Evolutionarily Conserved Complex Required for Efficient Homologous
  Recombination Repair - JBC (2011)


• Enzyme Stability of Microencapsulated Bifidobacterium animalis ssp. lactis Bb12 after Freeze Drying and during Storage in
  Low Water Activity at Room Temperature - Journal of Food Science (2011)


• Digitoxin and a synthetic monosaccharide analog
  inhibit cell viability in lung cancer cells - Toxicology and Applied
  Pharmacology (2011)


• Functional
  defect caused by the 4544G>A SNP in ABCC2: potential impact for drug
  cellular disposition - Pharmacogenetics and Genomics (2011)


• A
  Common Variant in TFB1M
  Is Associated with Reduced Insulin Secretion and Increased Future Risk of
  Type 2 Diabetes - Cell Metabolism (2011)


• Overexpression of the Endoplasmic Reticulum
  Chaperone BiP3 Regulates XA21-Mediated Innate Immunity in Rice
  – PLoS One (2010)


• Identification of gp96 as a Novel Target for
  Treatment of Autoimmune Disease in Mice - PLoS One (2010)


• Antimicrobial
  Mechanism of Action of Transferrins: Selective Inhibition of H⁺-ATPase - Antimicrob Agents Chemother.
  (2010)


• Modulation of UvrD
  Helicase Activity by Covalent DNA-Protein Cross-links - Biol Chem. (2010)


• Monoclonal
  antibodies against nucleoside triphosphate hydrolase-II can reduce the
  replication of Toxoplasma gondii - Parasitology (2010)


• Biochemical Dissection
  of the ATPase TraB, the VirB4 Homologue of theEscherichia coli pKM101
  Conjugation Machinery - J. Bacteriol. (2010)


• Identification of
  gp96 as a Novel Target for Treatment of Autoimmune Disease in Mice
  – PLoS One (2010)


• ATP-binding
  cassette-like transporters are involved in the transport of lignin
  precursors across plasma and vacuolar membranes - PNAS (2010)


• Arabidopsis stromal 70-kDa heat shock proteins
  are essential for chloroplast development - Planta (2010)


• Lipopeptide induces apoptosis in fungal cells by
  a mitochondria-dependent pathway - Peptides (2010)


• Modulation of a
  Schistosoma mansoni multidrug transporter by the antischistosomal drug
  praziquantel -
  The FASEB Journal (2010)


• Biosynthesis of (Bacterio)chlorophylls
  ATP-Dependent Transient Subunit Interaction And Electron Transfer Of Dark
  Operative Protochlorophyllide Oxireductase - JBC (2010)


• Stress-Dependent Coordination of Transcriptome
  and Translatome in Yeast - PLoS Biology (2009)


• Crystal Structure
  of a Novel Conformational State of the Flavivirus NS3 Protein:
  Implications for Polyprotein Processing and Viral Replication - J
  Virol (2009)


• ATP and MO25α
  Regulate the Conformational State of the STRADα Pseudokinase and
  Activation of the LKB1 Tumour Suppressor – PLoS Biol (2009)


• Apratoxin A
  Reversibly Inhibits the Secretory Pathway by Preventing Cotranslational
  Translocation -
  Molecular Pharmacology (2009)


• NS4A
  regulates the ATPase activity of the NS3 helicase: a novel cofactor role
  of the non-structural protein NS4A from West Nile virus - Journal of General Virology
  (2009)


• Evidence That Two ATP-Dependent (Lon) Proteases
  in Borrelia burgdorferi
  Serve Different Functions - PLoS Pathogens (2009)


• ATP and MO25α Regulate the Conformational State
  of the STRADα Pseudokinase and Activation of the LKB1 Tumour Suppressor
  - PLoS Biology (2009)


• Stress-Dependent Coordination of Transcriptome
  and Translatome in Yeast - PLoS Biology (2009)


• An Afg2/Spaf-Related Cdc48-like AAA ATPase
  Regulates the Stability and Activity of the C. elegans Aurora B Kinase AIR-2 - Developmental Cell (2008)


• Staphylococcus aureus HrtA Is an ATPase Required for Protection
  against Heme Toxicity and Prevention of a Transcriptional Heme Stress
  Response -
  Journal of Bacteriology (2008)


• ATP
  scavenging by the intracellular pathogen Porphyromonas gingivalis inhibits
  P2X₇-mediated host-cell apoptosis - Cell Microbiol. (2008)


• The Two-component NS2B-NS3 Proteinase Represses DNA
  Unwinding Activity of the West Nile Virus NS3 Helicase - JBC (2008)


• Functional characterization of the nucleotide
  binding domain of the Cryptosporidium parvum CpABC4 transporter: An iron–sulfur cluster
  transporter homolog -
  Molecular and Biochemical Parasitology (2008)


• Hippocampal
  levels of gamma-enolase, C-1-tetrahydrofolate synthase and serotransferrin
  fluctuate over the estrous cycle in the rat - Neuroscience (2008)


• Consumption of Soy Protein Isolate Modulates the
  Phosphorylation Status of Hepatic ATPase/ATP Synthase ß Protein and
  Increases ATPase Activity in Rats - J. Nutr. (2007)


• First identification and functional
  characterization of an immunogenic protein in unculturable haemotrophic
  Mycoplasmas (Mycoplasma suis HspA1) - FEMS Immunology & Medical
  Microbiology (2007)


• Identification,
  structure and mode of action of a new regulator of the Helicobacter pylori
  HP0525 ATPase -
  EMBO J. (2007)


• Proteome Profiling in
  the Rat Harderian Gland - J Proteome Res
  (2006)

GTPase Assay Kit

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Product

• Tubulin nucleotide status controls
  Sas-4-dependent pericentriolar material recruitment - Nature Cell Biology (2012)


• The Mitochondrial Phosphatase PGAM5 Functions at
  the Convergence Point of Multiple Necrotic Death Pathways
  - Cell (2012)


• Leucyl-tRNA Synthetase Is an Intracellular
  Leucine Sensor for the mTORC1-Signaling Pathway - Cell (2012)


• ArfGAP1 Is a GTPase Activating Protein for
  LRRK2: Reciprocal Regulation of ArfGAP1 by LRRK2 - Journal of Neuroscience (2012)


• GTPase Activity and Neuronal Toxicity of
  Parkinson's Disease–Associated LRRK2 Is Regulated by ArfGAP1
  - PLoS Genetics (2012)


• AIP1 Functions as Arf6-GAP to Negatively
  Regulate TLR4 Signaling - JBC (2010)


• GTPase activity plays a key role in the
  pathobiology of LRRK2 - PLoS Genetics (2010)


• Role of the
  G-protein and tyrosine kinase—Rho/ROK pathways in
  15-hydroxyeicosatetraenoic acid induced pulmonary vasoconstriction in
  hypoxic rats - J Biochem (2010)


• AIP1 Functions as Arf6-GAP to Negatively
  Regulate TLR4 Signaling - JBC (2010)


• Identification and Characterization of a
  Leucine-Rich Repeat Kinase 2 (LRRK2) Consensus Phosphorylation Motif
  - PLoS One (2010)


• Speckled-like Pattern in the Germinal Center
  (SLIP-GC), a Nuclear GTPase Expressed in Activation-induced
  Deaminase-expressing Lymphomas and Germinal Center B Cells
  - JBC (2009)


• High-level production and characterization of a
  G-protein coupled receptor signaling complex - FEBS Journal (2009)


• Structure of ERA in
  complex with the 3′ end of 16S rRNA: Implications for ribosome biogenesis
  - PNAS (2009)


• G Protein-Coupled Receptor-Promoted Trafficking
  of Gβ1γ2 Leads to AKT Activation at Endosomes via a Mechanism Mediated by
  Gβ1γ2-Rab11a Interaction – Molecular Biology of the Cell (2008)


• Characterization
  of Human GTPBP3, a GTP-Binding Protein Involved in Mitochondrial tRNA
  Modification - Molecular
  and Cellular Biology (2008)


• Comparative real-time PCR and enzyme analysis of
  selected gender-associated molecules in Schistosoma japonicum - Parasitology (2008)

PiColorlock Gold

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Product

• Kinetic Modelling of GlmU Reactions –
  Prioritization of Reaction for Therapeutic Application
  - PLoS One (2012)


• The unusual mycobacterial chaperonins: evidence
  for in vivo oligomerization and specialization of function
  - Molecular
  Microbiology (2012)


• Synthesis and Antibacterial Evaluation of a New
  Series of N-Alkyl-2-alkynyl/(E)-alkenyl-4-(1H)-quinolones
  - Molecules (2012)


• PPM1H Is a p27 Phosphatase Implicated in
  Trastuzumab Resistance - Cancer Discovery (2011)


• Sepsis impairs alveolar epithelial function by
  downregulating Na-K-ATPase pump - AJP - Lung Physiol (2011)


• Structural
  and functional insights into DNA-end processing by the archaeal HerA
  helicase–NurA nuclease complex - Nucleic Acids Research (2011)


• Sepsis
  impairs alveolar epithelial function by downregulating Na-K-ATPase pump
  - AJP - Lung Physiol (2011)


• FXYD Proteins Stabilize Na,K-ATPase,
  Amplification Of Specific Phosphatidylserine -Protein Interactions
  - JBC (2011)


• Structural
  and functional insights into DNA-end processing by the archaeal HerA
  helicase–NurA nuclease complex - Nucl Acids Res
  (2011)


• Stabilization
  of the α2 isoform of Na,K-ATPase by mutations in a phospholipid binding
  pocket - JBC (2011)


• Characterisation of multiple substrate-specific (d)ITP/(d)XTPase and
  modelling of deaminated purine nucleotide metabolism - MIMS EPrints (2011)


• Substrate uptake, phosphorus repression, and
  effect of seed culture on glycopeptide antibiotic production: Process
  model development and experimental validation - Biotechnology and
  Bioengineering (2010)


• Selectivity of Digitalis Glycosides for Isoforms
  of Human Na,K-ATPase
  - JBC (2010)


• ATP-dependent
  MurE ligase in Mycobacterium
  tuberculosis: Biochemical and structural characterisation -
  Tuberculosis (2010)


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