ATPase Activity Kit (Colorimetric)

Product Discontinued

ATPase Activity Kit (Colorimetric) Summary

• Description: This non radioactive colorimetric assay kit contains all the necessary reagents for measuring enzyme activity (everything included in the PiColorlock kit and more) and is ideal for high throughput drug screening.
  
  The kits contain PiColorlock, a non-radioactive, superior phosphate detection reagent.
  
  Key Features of PiColorLock:
  • Colorimetric Assay-competitor assays are radioactive.
  • Special additive ensures low backgrounds with acid-labile substrates.
  • Unparalleled stability of phosphate-dye complexes.
  • Reagent is compatible with almost any assay buffer.
  • No inhibition of color development by high concentrations of protein.
  • Stable reagent formulation - long shelf life.
  Colorimetric assays for ATPases are invariably based on the formation of colored complexes between an inorganic phosphate and a dye molecule under acidic conditions.
• Specificity: In vitro assay as reported in the literature (PMID: 21304972)
• Kit Type: Activity Kit (Colorimetric)
• Gene: DNAH8

Applications/Dilutions

• Dilutions:
  • In vitro assay

Packaging, Storage & Formulations

• Storage: Storage of components varies. See protocol for specific instructions.

Notes

This product is manufactured by Expedeon Inc. and distributed by Novus Biologicals.

Alternate Names for ATPase Activity Kit (Colorimetric)

• ATPase
• axonemal
• dynein, axonemal, heavy chain 8
• dynein, axonemal, heavy polypeptide 8
• FLJ25850
• FLJ36115
• FLJ36334
• hdhc9

Background

CATALYTIC ACTIVITY: ATP + H2O = ADP + phosphate. SUBCELLULAR LOCATION: Membrane; multi-pass membrane protein (By similarity). ALTERNATIVE PRODUCTS: 2 named isoforms produced by alternative splicing. SIMILARITY: Belongs to the cation transport ATPase (P-type) family. Type V subfamily.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Publications for ATPase Kit (601-0121)(65)

Publications using 601-0121

• Zadorozhny K, Sannino V, Belan O et al. Fanconi-Anemia-Associated Mutations Destabilize RAD51 Filaments and Impair Replication Fork Protection. Cell Rep. 2017-10-10 [PMID: 29020621]
• Granja AG, Tafalla C. Different IgM+ B cell subpopulations residing within the peritoneal cavity of vaccinated rainbow trout are differently regulated by BAFF. Fish Shellfish Immunol. 2017-10-05 [PMID: 28989090]
• Park SA, Hong BZ, Ha KC et al. Protein tyrosine phosphatase 1B is a mediator of cyclic ADP ribose-induced Ca2+ signaling in ventricular myocytes. Exp Mol Med. 2017-06-02 [PMID: 28572573]
• Bittencourt IA, Serpeloni M, Hiraiwa PM et al. Dissecting biochemical peculiarities of the ATPase activity of TcSub2, a component of the mRNA export pathway in Trypanosoma cruzi . Int J Biol Macromol. 2017-01-01 [PMID: 28212935]
• Huerta-Uribe A, Marjenberg ZR, Yamaguchi N et al. Identification and Characterization of Novel Compounds Blocking Shiga Toxin Expression in Escherichia coli O157:H7. Front Microbiol. 2016-11-30 [PMID: 27965652]
• Martino L, Holland L, Christodoulou E et al. The Biophysical Characterisation and SAXS Analysis of Human NLRP1 Uncover a New Level of Complexity of NLR Proteins. PLoS One. 2016-10-11 [PMID: 27727326]
• Sheng Y, Yang X, Lian Y et al. Characterization of a cadmium resistance Lactococcus lactissubsp. lactis strain by antioxidant assays and proteome profiles methods. Environ Toxicol Pharmacol. 2016-01-01 [PMID: 27522548]
• Kristensen ML, Kierulf-Lassen C, Nielsen PM et al. Remote ischemic perconditioning attenuates ischemia/reperfusion-induced downregulation of AQP2 in rat kidney. Physiol Rep. 2016-01-01 [PMID: 27405971]
• Wambach JA, Yang P, Wegner DJ et al. Functional Characterization of ABCA3 Mutations from Infants with Respiratory Distress Syndrome. Am J Respir Cell Mol Biol. 2016-01-01 [PMID: 27374344]
• Wang Y, Sun Y, Zhang Y et al. Antifungal Activity and Biochemical Response of Cuminic Acid against Phytophthora capsici Leonian. Molecules. 2016-06-11 [PMID: 27294911]
• Dellas N, Snyder JC, Dills M et al. Structure-Based Mutagenesis of Sulfolobus Turreted Icosahedral Virus B204 Reveals Essential Residues in the Virion-Associated DNA-Packaging ATPase. J Virol. 2015-12-23 [PMID: 26699645]
• Huang SW, Liu HP, Chen JK et al. Dual ATPase and GTPase activity of the replication-associated protein (Rep) of beak and feather disease virus. Virus Res. 2016-02-02 [PMID: 26657880]
• Ma ZX, Leng YJ, Chen GX et al. The THERMOSENSITIVE MALE STERILE 1 Interacts with the BiPs via DnaJ Domain and Stimulates Their ATPase Enzyme Activities in Arabidopsis. PLoS One. 2015-07-17 [PMID: 26186593]
• Ono H, Takashima A, Hirata H et al. The MinD homolog FlhG regulates the synthesis of the single polar flagellum of Vibrio alginolyticus. Mol Microbiol. 2015-01-01 [PMID: 26112286]
• Zhang J, He S, Wang Y et al. Herpesviral G Protein-Coupled Receptors Activate NFAT to Induce Tumor Formation via Inhibiting the SERCA Calcium ATPase. PLoS Pathog. 2015-03-26 [PMID: 25811856]
• Fishburn J, Tomko E, Galburt E, Hahn S. Double-stranded DNA translocase activity of transcription factor TFIIH and the mechanism of RNA polymerase II open complex formation . Proc Natl Acad Sci U S A. 2015-03-31 [PMID: 25775526]
• Shao W, Zhang Y, Ren W, Chen C2. Physiological and biochemical characteristics of laboratory induced mutants of Botrytis cinerea with resistance to fluazinam. Pestic Biochem Physiol. 2015-01-01 [PMID: 25619907]
• Torrente MP, Castellano LM, Shorter J Suramin Inhibits Hsp104 ATPase and Disaggregase Activity. PLoS One. 2014-10-09 [PMID: 25299406]
• Sarraf NS, Shi R, McDonald L et al. Suramin Inhibits Hsp104 ATPase and Disaggregase Activity. PLoS One 2014-01-01 [PMID: 24945826]
• Xiao X, Yang J, Li R et al. A helminth cathelicidin-like protein suppresses antigen processing and presentation in macrophages via inhibition of lysosomal vATPase. FASEB J. 2014-01-01 [PMID: 24391795]
• Matos M, Augusto E, Agostinho P et al. Antagonistic Interaction between Adenosine A2A Receptors and Na+/K+-ATPase-alpha 2 Controlling Glutamate Uptake in Astrocytes. J Neurosci 2013-01-01 [PMID: 24259572]
• Luo H, Xu HQ, Chen X et al. Potent in vitro Interference of Fleroxacin in DNA-binding, Unwinding and ATPase Activities of Bloom Helicase. Biomed Environ Sci. 2013-01-01 [PMID: 23534463]
• Bruchmann A, Roller C, Walther TV et al. Bcl-2 associated athanogene 5 Bag5is overexpressed in prostate cancer and inhibits ER-stress induced apoptosis. BMC Cancer 2013-01-01 [PMID: 23448667]
• Ronchi D, Di Fonzo A, Lin W et al. Mutations in DNA2 Link Progressive Myopathy to Mitochondrial DNA Instability. Am J Hum Genet. 2013-01-01 [PMID: 23352259]
• Choi CH1, Spooner R, DeGuzman J et al. P. gingivalis-Nucleoside-diphosphate-kinase Inhibits ATP-Induced Reactive-Oxygen-Species via P2X7 Receptor / NADPH-Oxidase Signaling and Contributes to Persistence. Cell Microbiol. 2013-01-01 [PMID: 23241000] (EnzAct, Bacteria)
  
  Details:
  Porphyromonas gingivalis
• Robinson MW, Alvarado R, To J et al. Physiological and biochemical characteristics of laboratory induced mutants of Botrytis cinerea with resistance to fluazinam. FASEB J. 2014-01-01 [PMID: 22872675]
• Baker N, Zhang G, You Y, Tuan R. Caveolin-1 regulates proliferation and osteogenic differentiation of human mesenchymal stem cells. J Cell Biochem 2012-01-01 [PMID: 22807396]
• Podhajska A, Musso A, Trancikova A et al. Common Pathogenic Effects of Missense Mutations in the P-Type ATPase ATP13A2 PARK9 Associated with Early-Onset Parkinsonism. PLoS One 2012-01-01 [PMID: 22768177]
• Wallden K, Williams R, Yan J et al. Structure of the VirB4 ATPase, alone and bound to the core complex of a type IV secretion system. PNAS 2012-01-01 [PMID: 22745169]
• Bartoszewska M, Williams C, Kikhney A et al. Peroxisomal proteostasis involves a Lon family protein that functions as protease and chaperone. J Biol Chem. [PMID: 22733816] (EnzAct, Bacteria)
  
  Details:
  Penicillium chrysogenum
• Firestone AJ, Weinger JS, Maldonado M et al. Small-molecule inhibitors of the AAA+ ATPase motor cytoplasmic dynein Nature 2012-03-01 [PMID: 22425997]
• Tan LJ, Saijo M, Kuraoka I et al. Xeroderma pigmentosum group F protein binds to Eg5 and is required for proper mitosis implications for XP-F and XFE. Genes Cells 2012-01-01 [PMID: 22353549]
• Elbaz HA, Stueckle TA, Wang HY et al. Digitoxin and a synthetic monosaccharide analog inhibit cell viability in lung cancer cells. Toxicol Appl Pharmacol 2011-01-01 [PMID: 22037315]
• Elens L, Tyteca D, Panin N et al. Functional defect caused by the 4544G>A SNP in ABCC2 potential impact for drug cellular disposition. Pharmacogenet Genomics. 2011-01-01 [PMID: 22027652]
• Liu T, Wan L, Wu Y, Chen J et al. hSWS1 SWSAP1 Is an Evolutionarily Conserved Complex Required for Efficient Homologous Recombination Repair. J Biol Chem. 2011-12-02 [PMID: 21965664]
• Dianawati D, Shah NP. Enzyme Stability of Microencapsulated Bifidobacterium animalis ssp. lactis Bb12 after Freeze Drying and during Storage in Low Water Activity at Room Temperature. J Food Sci 2011-01-01 [PMID: 21696390]
• Koeck T, Olsson AH, Nitert MD et al. A Common Variant in TFB1M Is Associated with Reduced Insulin Secretion and Increased Future Risk of Type 2 Diabetes. Cell Metab 2011-01-01 [PMID: 21195351]
• Miao YC, Liu CJ. ATP-binding cassette-like transporters are involved in the transport of lignin precursors across plasma and vacuolar membranes. PNAS 2010-01-01 [PMID: 21149736]
• Qi G, Zhu F, Du P et al. Lipopeptide induces apoptosis in fungal cells by a mitochondria-dependent pathway. Peptides 2010-01-01 [PMID: 20713103]
• Andres MT, Fierro JF. Antimicrobial Mechanism of Action of Transferrins: Selective Inhibition of H+-ATPase. Antimicrob Agents Chemother. 2010-01-01 [PMID: 20625147]
• Latijnhouwers M, Xu XM, Moller SG. Arabidopsis stromal 70-kDa heat shock proteins are essential for chloroplast development. Planta 2010-01-01 [PMID: 20506024]
• Kumari A, Minko IG, Smith RL et al. Modulation of UvrD Helicase Activity by Covalent DNA-Protein Cross-links. J Biol Chem. [PMID: 20444702] (EnzAct, Virus)
  
  Details:
  West Nile Virus
• Han JM, Kwon NH, Lee JY et al. Identification of gp96 as a Novel Target for Treatment of Autoimmune Disease in Mice. PLoS One 2010-01-01 [PMID: 20352117]
• Park CJ, Bart R, Chern M et al. Overexpression of the Endoplasmic Reticulum Chaperone BiP3 Regulates XA21-Mediated Innate Immunity in Rice. PLoS One. 2010-02-17 [PMID: 20174657]
• Durand E, Oomen C, Waksman G. Biochemical Dissection of the ATPase TraB, the VirB4 Homologue of theEscherichia coli pKM101 Conjugation Machinery. J. Bacteriol. 2010-01-01 [PMID: 20172994]
• Chen X, Chern M, Canlas PE et al. Overexpression of the Endoplasmic Reticulum Chaperone BiP3 Regulates XA21-Mediated Innate Immunity in Rice - PLoS One. J Biol Chem 2010-01-01 [PMID: 20118235]
• Brocker MJ, Watzlich D, Saggu M et al. Biosynthesis of Bacteriochlorophylls ATP-Dependent Transient Subunit Interaction And Electron Transfer Of Dark Operative Protochlorophyllide Oxireductase. J Biol Chem 2010-01-01 [PMID: 20075073]
• Tan F, Hu X, Pan CW et al. Monoclonal antibodies against nucleoside triphosphate hydrolase-II can reduce the replication of Toxoplasma gondii. Parasitology 2010-01-01 [PMID: 20056166]
• Coleman JL, Katona LI, Kuhlow C et al. Evidence That Two ATP-Dependent (Lon) Proteases in Borrelia burgdorferi Serve Different Functions. PLoS Pathog. 2009-01-01 [PMID: 19956677]
• Assenberg R, Mastrangelo E, Walter TS et al. Crystal Structure of a Novel Conformational State of the Flavivirus NS3 Protein : Implications for Polyprotein Processing and Viral Replication. J Virol 2009-01-01 [PMID: 19793813]
• Kasinathan RS, Goronga T, Messerli SM et al. Modulation of a Schistosoma mansoni multidrug transporter by the antischistosomal drug praziquantel. FASEB J. 2010-01-01 [PMID: 19726755]
• Zeqiraj E, Filippi BM, Goldie S et al. ATP and MO25a Regulate the Conformational State of the STRADa Pseudokinase and Activation of the LKB1 Tumour Suppressor. PLoS Biol, 7 (6). 2009-01-01 [PMID: 19513107]
• Shiryaev SA, Chernov AV, Aleshin AE et al. NS4A regulates the ATPase activity of the NS3 helicase a novel cofactor role of the non-structural protein NS4A from West Nile virus. J Gen Virol. 2009-01-01 [PMID: 19474250] (EnzAct, E. coli)
  
  Details:
  Staphylococcus aureus
• Benitez AJ, Arrowood MJ, Mead JR. Functional characterization of the nucleotide binding domain of the Cryptosporidium parvum CpABC4 transporter :An iron-sulfur cluster transporter homolog. Mol Biochem Parasitol 2009-01-01 [PMID: 19428657]
• Halbeisen RE, Gerber AP. Stress-Dependent Coordination of Transcriptome and Translatome in Yeast. PLoS Biol. 2009-01-01 [PMID: 19419242]
• Liu Y, Law BK, Luesch H. Apratoxin A Reversibly Inhibits the Secretory Pathway by Preventing Cotranslational Translocation. Mol Pharm. 2009-01-01 [PMID: 19403701]
• Heallen TR, Adams HP, Furuta T et al. An Afg2/Spaf-Related Cdc48-like AAA ATPase Regulates the Stability and Activity of the C. elegans Aurora B Kinase AIR-2. Dev Cell 2008-01-01 [PMID: 18854144]
• Diao WF, Afjehi-Sadat L, Chen WQ et al. Hippocampal levels of gamma-enolase, C-1-tetrahydrofolate synthase and serotransferrin fluctuate over the estrous cycle in the rat. Neuroscience 2008-01-01 [PMID: 18511206]
• Stauff DL, Bagaley D, Torres VJ et al. The Two-component NS2B-NS3 Proteinase Represses DNA Unwinding Activity of the West Nile Virus NS3 Helicase. JBC 2008-01-01 [PMID: 18442976]
• Stauff DL, Bagaley D, Torres VJ et al. Staphylococcus Aureus HrtA Is an ATPase Required for Protection Against Heme Toxicity and Prevention of a Transcriptional Heme Stress Response . J Bacteriol. 2008-01-01 [PMID: 18326576] (EnzAct, Bacteria)
  
  Details:
  Staphylococcus aureus
• Yilmaz O, Yao L, Maeda K et al. ATP scavenging by the intracellular pathogen Porphyromonas gingivalis inhibits P2X7-mediated host-cell apoptosis. Cell Microbiol , 10 (4) pp 863-875. 2008-01-01 [PMID: 18005240]
• Hare S, Fischer W, Williams R et al. Identification, Structure and Mode of Action of a New Regulator of the Helicobacter Pylori HP0525 ATPase. EMBO Journal , 26 pp 4926-4934. 2007-01-01 [PMID: 17972918]
• Mei J, Wood C, L'abbe MR, Gilani GS, Cooke GM, Curran IH, Xiao CW. Consumption of soy protein isolate modulates the phosphorylation status of hepatic ATPase/ATP synthase beta protein and increases ATPase activity in rats. J. Nutr. 137: 2029-2035. 2007-01-01 [PMID: 17709438]
• Hoelzle LE, Hoelzle K, Harder A et al. First identification and functional characterization of an immunogenic protein in unculturable haemotrophic Mycoplasmas Mycoplasma suis HspA1. FEMS Immunol Med Microbiol 2007-01-01 [PMID: 17328755]
• Yang JW, Afjehi-Sadat L, Gelpi E et al. Proteome Profiling in the Rat Harderian Gland. J Proteome Res 2006-01-01 [PMID: 16823983]

FAQs for ATPase Kit (601-0121). (Showing 1 - 5 of 5 FAQs).

1. I have phosphate in my enzyme. What can I do?
   • You can dialyse or desalt the enzyme into a phosphate-free buffer. Alternatively, you can use a special resin (PiBind) to remove the phosphate.
2. I have 5% DMSO in my assay. Can I use PiColorLock Gold?
   • Yes, the reagent is designed for drug screening work and other situations that require DMSO.
3. I have a high background in my ATPase assay and I definitely do not have free phosphate in my sample
   • This is almost always due to inadequate mixing of the special stabilizer with the sample and detection reagent. Make sure the stabilizer is pipetted up and down several times to ensure thorough mixing.
4. I would like to measure the conversion of pyrophosphate to phosphate. Can I use the PiColorLock Gold Phosphate Detection System for this purpose?
   • Yes, only the phosphate will give a signal; pyrophosphate will not.
5. What is the ratio between ATP and ATPase in terms of molar, which may be appropriate for this assay?
   • There is no real definitive answer to this, as it depends on a variety of different things. The simple answer however is that there needs to be a huge excess of substrate. There needs to be enough substrate so this does not run out over the course of the assay, and stays in excess even as the enzyme reacts here, and increases the concentration of products.

Bioinformatics

• Gene Symbol: DNAH8