Alkaline phosphatase assay kit is a highly sensitive, simple, direct and htsready colorimetric assay designed to measure alp activity in serum and biological samples. Need pp2a immunoprecipitation phosphatase assay kit for research. Biochemphysiol actions acid phosphatase is one of the acid hydrolases that normally reside in lysosomes. The kit contains all the reagents required for a fast and simple acid phosphatase detection. The acid phosphatase assay kit is designed for the detection of acid phosphatase activity in whole cell and tissue extracts, column fractions, and purified enzyme preparations. Immunoprecipitation and malachite green phosphatase assay. Modulation of protein phosphatase 2a pp2a activity alters. Pp2a immunoprecipitation phosphatase assay kit upstate, temecula, ca was used to measure phosphate release as an index of phosphatase activity according to the manufacturers instructions.
However, this is an enzymebased kit so as long as your mouse myoblast cell line expresses alp ab83371 should work. Material safety data sheet or sds for pp2a immunoprecipitation phosphatase assay kit 173 from merck for download or viewing in the browser. Cellular protein phosphatase 2a pp2a activity was assayed using a pp2a immunoprecipitation phosphatase assay kit upstate. Modulation of protein phosphatase 2a pp2a activity alters androgenindependent growth of prostate cancer cells. Pdf phosphatase pp2a is requisite for the function of. Cells were washed in tbs, then lysed on ice in phosphatase assay buffer 20mm imidazolehcl, 2mm edta, 2mm egta, 0. Acid phosphatase assay kit kit sufficient for 1,000 assays. Protein phosphatase 2a pp2a and the related phosphatases. Highthroughput ready colorimetric assay to measure acid phosphatase ap activity in cells, tissues, serum, plasma, urine, semen and cell culture media. Alkaline phosphatase assay kit fluorometric has only been characterized on human sample. Phophatases hydrolyze a variety of organic phosphate monoesters to alcohols and inorganic phosphate, and have quite a few subfamilies.
Bioassay systems alkaline phosphatase assay kit alkaline. The rediplate 96 enzchek serinethreonine phosphatase assay kit provides a fast, simple and direct fluorescencebased assay for detecting serinethreonine phosphatases and their corresponding modulators and inhibitors. Biosassay systems pnpp phosphatase popn500 assay kit involves adding a single reagent to the sample the reaction yields paranitrophenol, which turns a yellow color under alkaline conditions and can be measured at 405 nm. Acid phosphatase assay kit cs0740 technical bulletin. I have treated my cells with increasing concentrations of a pp2a activator for 24h. Supernatant was then assayed with the pp2a immunoprecipitation phosphatase assay kit millipore, ma, usa. The cells were maintained in dmem supplemented with 10% fcs and penicillin g 100 unitsmlstreptomycin. Promega manufacturing and delivery systems continue to be fully operational during the covid19 outbreak. The activity of pp2a was analyzed using a nonradioactive pp2a immunoprecipitation phosphatase assay kit upstate biotechnology according to. Acid phosphatase activity colorimetric assay kit k411.
Bioassay systems quantichrom alkaline phosphatase assay kit is designed to measure alp activity directly in biological samples without pretreatment. Serinethreonine phosphatase assay system technical bulletin. It employs a convenient 96well microtiterplate format with all. Protein phosphatase type 2a pp2a is an essential protein serinethreonine phosphatase that is conserved in all eukaryotes. Alkaline phosphatase activity colorimetric assay kit.
The target phosphatase is bound by the capture antibody. Pp2a antibody for chromatin immunoprecipitation chip, immunoprecipitation ip. Rediplate 96 enzchek serinethreonine phosphatase assay kit. This kit is optimized to detect protein phosphatase activity in biological samples or to screen for. How to improve phosphatase activity of pp2a using the pp2a. Pp2a is a key enzyme within various signal transduction pathways as it regulates fundamental cellular activities such as dna replication, transcription, translation, metabolism, cell cycle progression, cell division, apoptosis and development. Pp2a activity was measured in cycling extracts using the pp2a immunoprecipitation phosphatase assay kit upstate. For acid phosphatase assay kit ab83370, the ph of the buffer prevents alkaline phosphatase from working alp can only function in an alkaline ph whereas ap works in an acidic ph so you only measure ap activity. The assay works with protein phosphatase 1 pp1, pp2a, pp2b, and pp2c. After immunoprecipitation, the activity of the pp2a was assessed by the release of phosphate from a chemically synthesized phosphopeptide over a period of.
Protein phosphatase 1 and protein phosphatase 2a are two of the. Though, optimization may be necessary since we have no data on murine system. This system determines the amount of free phosphate generated in a reaction by measuring the absorbance of a molybdate. Phosphatase assay kit emd millipore, temecula, ca following the. Protein phosphatase 2a pp2a is a major eukaryotic serinethreonine phosphatase that is highly conserved from yeast to human. A protocol for fast, convenient measuring of protein serinethreonine. Bioassay systems alkaline phosphatase assay kit manufacturer. Pp2a immunoprecipitation phosphatase assay kit was purchased from merck millipore. Unlike other microplate assays, this kit provides the necessary reagents predispensed into a 96well microplate. I am using the pp2a immunoprecipitation phosphatase assay kit from millipore. Rbc specializes in custom assay conditions, high quality reproducible data, and outstanding service. Assay kit colorimetric ab9464 phosphatase activity. Proteins were immunoprecipitated overnight with anti pp2a antibodies and protein gagarose beads.
Phosphatase pp2a is essential for th17 differentiation. Phpt1 is an edtainsensitive phosphohistidine phosphatase that catalyzes the dephosphorylation of phosphopeptide i ek et al. They are an important class of enzymes that are involved in cellular signal transduction, cell metabolism and other biological activities. Briefly, protein extracts were immunoprecipitated with a pp2a c specific antibody clone. Highthroughput ready colorimetric assay to measure alkaline phosphatase alp activity in cells, serum, plasma etc. Protein phosphatase 2a pp2a holoenzymes regulate death. Pp2a immunoprecipitation phosphatase assay kit msds 17. Pp2a activity was determined using pp2a immunoprecipitation phosphatase assay kit according to the manufacturers instructions.
Proteasephosphatase assay kits, kits, products creative. The alkaline phosphatase assay kit is designed to measure alp activity directly in biological samples without pretreatment. The nonradioactive serinethreonine phosphatase assay system provides a fast, convenient and flexible method for measuring protein serinethreonine phosphatase activity. Targeting set to restore pp2a activity disrupts an. A novel assay for protein phosphatase 2a pp2a complexes. Dapk substrate peptide was from tocris ellisville, mo. Alkaline phosphatase assay kit bioassay systems 8777823888. Arctigenin inhibits triplenegative breast cancers by. Find msds or sds, a coa, data sheets and more information. Lysates of transfected cells were incubated with 50 l50% slurry of anti. A colorimetric protein phosphatase inhibition assay ppi assay, a commercial enzymelinked immunosorbent assay elisa test and different hplc methods using uv detection were compared for the detection of cyanobacterial hepatotoxins, microcystins mcyst and nodularin.
Protein phosphatase 2a pp2a and the related phosphatases pp4 and pp6. Protein phosphatase 2 pp2, also known as pp2a, is an enzyme that in humans is encoded by the ppp2ca gene. The phosphatase assay kit is designed to measure the activity of phosphatases in biological samples and to screen for agonists and inhibitors of phosphatases. Protein phosphatase 2a pp2a is a major serinethreonine protein phosphatase that regulates multiple cellular processes 1. Pp2a immunoprecipitation phosphatase assay kit was from millipore temecula, ca. The kit contains a standard solution and a control enzyme. Protein phosphatase 1 an overview sciencedirect topics. Serthr phosphatase assay kit 1 krptirr sigmaaldrich.
It is a marker for the identification of lysosomes in subcellular fractionations. The improved method utilizes pnitrophenyl phosphate that. Pp2a immunoprecipitation phosphatase assay kit was from upstate biotechnology lake placid, ny. The phosphatase assay kit uses paranitrophenyl phosphate pnpp, a chromogenic substrate for most phosphatases, including alkaline. Detection of microcystins with protein phosphatase inhibition. Find and purchase phosphatase assay kits products at invitrogen life science technologies. Pp2a immunoprecipitation phosphatase assay kit upstate, temecula, ca, usa was used to measure phosphate release as an index of phosphatase activity according to the manufacturers instructions.
Pp2a is able to participate in such a wide array of processes because of its. The pp2a heterotrimeric protein phosphatase is ubiquitously expressed, accounting for a large fraction of phosphatase activity in eukaryotic cells. A new regulatory mechanism of protein phosphatase 2a. Cd25 and protein phosphatase 2a cooperate to enhance il2r. Can be used to detect purified pp2a or pp2a from a cellular lysate. Immunoprecipitation phosphatase assay pp1 and pp2a phosphatase activity were measured with cell lysates 500 lg prepared from the whole cell lysates using an immunoprecipitation phosphatase assay kit millipore, billerica, ma, usa, following the manufacturers protocol. Pp2a activity has been functionally linked to a variety of cellular processes including cell cycle progression, dna replication, transcription, rna splicing, and translation 9, 17, 35.
Pp2a c enzymatic activity was assessed using the pp2a immunoprecipitation phosphatase. Carfilzomib induces leukaemia cell apoptosis via inhibiting. Lipofectamine 2000 sirna transfection reagent was from invitrogen. Principle of the assay abcam s cellular calcineurin phosphatase activity assay kit colorimetric ab9464 is a complete colorimetric assay kit for measuring cellular calcineurin pp2b phosphatase activity. Phosphatase in regulating 1433 release from cdc25 to control mitosis. This alkaline phosphatase assay kit is a highly sensitive, simple, direct and htsready colorimetric assay designed to measure alp activity in serum and biological samples specificity this kit can detect all forms of alkaline phosphatase intestinal, placental, germ cell, liverbonekidney. Its serinethreonine phosphatase activity has a broad substrate specificity and diverse cellular functions. Pp2a immunoprecipitation phosphatase assay kit from. Pp2a is involved in the regulation of a number of signaling pathways, and it is implicated in the negative. The promoter activity was determined by using the dualluciferase reporter assay kit promega, madison, wi, usa according to. Listed are elisa kits for the detection of pp2a, an alias name of protein phosphatase 2 phosphatase activator. Phosphatase pp2a is requisite for the function of regulatory t cells. Brain phosphatase activity was determined using the serthr phosphatase assay kit from millipore temecula, ca according to the manufacturers instructions.
With the largest functional kinase activity assay panel in the industry, rbc provides kinase, epigenetic, and other profiling services to over 750 companies worldwide. Regulation of protein phosphatase 2a by arpp 16 and mast3. Milliporesigma pp2a immunoprecipitation phosphatase assay kit pp2a immunoprecipitation phosphatase assay kit life sciences. The improved method utilizes pnitrophenyl phosphate that is hydrolyzed by alp into a yellow colored product maximal absorbance at 405nm. Pp2a immunoprecipitation phosphatase assay kit pp2a immunoprecipitation phosphatase assay kit sds safety data sheet for pp2a immunoprecipitation phosphatase assay kit 173. Acid phosphatase is one of the acid hydrolases that normally reside in lysosomes.
Alkaline phosphatase assay kit colorimetric ab83369 is a highly sensitive, simple, direct and htsready colorimetric assay designed to measure alkaline phosphatase alp activity in serum and other mammalian samples. Immunocomplexes were washed three times in lysis buffer without phosphatase inhibitors and two times in pp2a reaction buffer50 mm tris. Pp2a elisa kits the elisa enzymelinked immunosorbent assay is a widely used application for detecting and quantifying proteins and antigens from various samples. The assay utilizes paranitrophenyl phosphate pnpp as a chromogenic substrate for the enzyme. Phosphatase assay kit 500 tests bioassay systems 877. It is a classical marker for the identification of lysosomes in subcellular fractionations. Pp2a inhibition from lb100 therapy enhances daunorubicin. Pp2a immunoprecipitation phosphatase assay kit 173. Protein phosphatase 2a pp2a journal of biological chemistry. Locate proteins, assay kits, reagents, custom services. Pp2a antibody for chromatin immunoprecipitation chip. Study of protein phosphatase 2a pp2a activity in lpsinduced. Simply reconstitute the fluorogenic substrate in the assay.
Each kit contains a sufficient amount of substrate for assays, using a reaction volume of 100 l per assay. Overexpression of cip2a is associated with poor prognosis in. Colorimetric immunoprotein phosphatase inhibition assay. About this assay caymans acid phosphatase colorimetric assay kit provides a convenient method for detecting total ap activity in plasma, serum, urine, and semen. Kits are available in 2, 5, and 1596well plate pack sizes. Pp2a immunoprecipitation phosphatase assay kit upstate, temecula, ca, usa was used to measure phosphate release as an index of phosphatase activity, according to the. Pp2a immunoprecipitation phosphatase assay kit can be used to detect purified pp2a or pp2a from a cellular lysate. The human protein, encoded by the gene ptpa, is 358 amino acid residues long and has a mass of 40,668 daltons.
Milliporesigma pp2a immunoprecipitation phosphatase assay kit. The results are expressed as foldchange of the control, which are assigned a. Study of protein phosphatase 2a pp2a activity in lps. Protein phosphatase 2a pp2a is one of the most abundant intracellular. Milliporesigma pp2a immunoprecipitation phosphatase assay kit utilizes malachite green and contains components that have been matched to optimize sensitivity and range of detection manufacturer. Find and compare commercial and governmental sources for immunological and biological products using the linscotts directory search engine. Our teams are in regular contact with suppliers and distributors worldwide and are taking all steps necessary to address both demands for diagnostic tools and reliable delivery of all products as quickly as possible. We addressed the problem of specificity in the analysis of protein phosphatase inhibitors by combining immunoassaybased detection of the toxins with a colorimetric protein phosphatase inhibition system in a single assay, designated the colorimetric immunoprotein phosphatase inhibition assay. Fugene 6 transfection reagent was purchased from roche diagnostics. Pp2a protein phosphatase 2a beta subunit is the catalytic subunit of protein phosphatase 2a. Alkaline phosphatase assay kit colorimetric ka0817.
Modulation of protein phosphatase 2a pp2a activity. The kit is perfect for the continuous assay of prostatic acid phosphatase, protein phosphatase 1 or almost any other phosphatase that can be assayed with nonproteinbased substrates such as mup or 4nitrophenyl phosphate pnpp. The phosphatase activity of pp2a were assessed using pp2a immunoprecipitation phosphatase assay kit millipore, billerica, ma, usa as previously describe. Pp2a immunoprecipitation phosphatase assay kit sds safety data sheet for pp2a immunoprecipitation phosphatase assay kit 173. Principle of the assay simple, direct and automationready procedures for measuring alp activity in serum are becoming popular in research and drug discovery.
The acid phosphatase assay kit colorimetric is a high sensitivity, simple, direct and htsready colorimetric assay designed to measure ap activity in serum and other. Targetspecific elisa kits are available from a variety of manufacturers and can help streamline your immunodetection experiments. Elevated prostatic acid phosphatase levels may indicate the presence of prostate cancer and elevated tartrateresistant acid phosphatase levels may indicate the bone disease. Enzcheck serthr phosphatase assay kit was purchased from molecular. The captured phosphatase liberates phosphate from the phosphopeptide substrate and the free phosphate is measured by a change in color of malachite green.
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