Genomes and Genes
phospho specific antibodies
Summary: Antibodies directed against immunogen-coupled phosphorylated PEPTIDES corresponding to amino acids surrounding the PHOSPHORYLATION site. They are used to study proteins involved in SIGNAL TRANSDUCTION pathways. (From Methods Mol Biol 2000; 99:177-89)
- Soulsby M, Wojcikiewicz R. Calcium mobilization via type III inositol 1,4,5-trisphosphate receptors is not altered by PKA-mediated phosphorylation of serines 916, 934, and 1832. Cell Calcium. 2007;42:261-70 pubmed..Overall, these data suggest that modification of Ca2+ flux may not be the primary effect of IP3R3 phosphorylation by PKA. ..
- Diks S, Peppelenbosch M. Single cell proteomics for personalised medicine. Trends Mol Med. 2004;10:574-7 pubmed..Thus, single cell phosphoproteomic techniques might be superior to other proteomic approaches for the molecular diagnosis of disease and instrumental for the development of personalised medicine. ..
- Lemaire M, Froment C, Boutros R, Mondesert O, Nebreda A, Monsarrat B, et al. CDC25B phosphorylation by p38 and MK-2. Cell Cycle. 2006;5:1649-53 pubmed..Since most of these sites are also phosphorylated by several other kinases, our observations highlight the difficulty in characterizing and understanding in vivo phosphorylation patterns. ..
- Lopez Bergami P, Habelhah H, Bhoumik A, Zhang W, Wang L, RONAI Z. RACK1 mediates activation of JNK by protein kinase C [corrected]. Mol Cell. 2005;19:309-20 pubmed..In finding the role of RACK1 in activation of JNK by PKC, our study also highlights the nature of crosstalk between these two signal-transduction pathways. ..
- Padmanabhan J. Detection of histone H3 phosphorylation in cultured cells and tissue sections by immunostaining. Methods Mol Biol. 2009;523:311-22 pubmed publisher..J Neurochem 90:1117-1131, 2004; Crosio et al., J Cell Sci 116:4905-4914, 2003). Co-staining with a cell-specific antibody will allow us to determine the type of cells that show activation of histone phosphorylation in the brain. ..
- Roldan J, Brey R. Neurologic manifestations of the antiphospholipid syndrome. Curr Rheumatol Rep. 2007;9:109-15 pubmed
- Raska C, Parker C, Sunnarborg S, Pope R, Lee D, Glish G, et al. Rapid and sensitive identification of epitope-containing peptides by direct matrix-assisted laser desorption/ionization tandem mass spectrometry of peptides affinity-bound to antibody beads. J Am Soc Mass Spectrom. 2003;14:1076-85 pubmed..The first application of this method was with anti-FLAG-tag affinity beads, where direct MALDI MS/MS was used to determine an unexpected enzymatic cleavage site on a growth factor protein. ..
- Schulz K, Danna E, Krutzik P, Nolan G. Single-cell phospho-protein analysis by flow cytometry. Curr Protoc Immunol. 2007;Chapter 8:Unit 8.17 pubmed publisher..This technique has broad applications to take standard biochemical analysis into primary cells to garner valuable information about signaling events in physiologic settings. ..
- Tarapore P, Shinmura K, Suzuki H, Tokuyama Y, Kim S, Mayeda A, et al. Thr199 phosphorylation targets nucleophosmin to nuclear speckles and represses pre-mRNA processing. FEBS Lett. 2006;580:399-409 pubmed..These findings indicate the involvement of NPM in the regulation of pre-mRNA processing, and its activity is controlled by CDK2-mediated phosphorylation on Thr(199). ..
- Ho M, Hoang H, Lee K, Liu N, Macrae T, Montes L, et al. Ectophosphorylation of CD36 regulates cytoadherence of Plasmodium falciparum to microvascular endothelium under flow conditions. Infect Immun. 2005;73:8179-87 pubmed..Collectively, these results are consistent with a critical role for CD36 dephosphorylation through Src family kinase activation in regulating IRBC adhesion to vascular endothelium. ..
- Zheng F, Wu R, Smith C, O Malley B. Rapid estrogen-induced phosphorylation of the SRC-3 coactivator occurs in an extranuclear complex containing estrogen receptor. Mol Cell Biol. 2005;25:8273-84 pubmed..Our results provide evidence for an early nongenomic action of ER on SRC-3 that supports the well-established downstream genomic roles of estrogen and coactivators. ..
- Xie S, Martin S, Guillot P, Bentley D, Pombo A. Splicing speckles are not reservoirs of RNA polymerase II, but contain an inactive form, phosphorylated on serine2 residues of the C-terminal domain. Mol Biol Cell. 2006;17:1723-33 pubmed..We find that paraspeckles are transcriptionally inactive but contain polymerase II, which remains stably associated upon transcriptional inhibition, when paraspeckles reorganize around nucleoli in the form of caps...
- Khanna K, Gatei M, Tribbick G. Phospho-specific antibodies as a tool to study in vivo regulation of BRCA1 after DNA damage. Methods Mol Med. 2006;120:441-52 pubmed..These antibodies can be used to detect a particular phosphorylation event in vivo using Western blotting, immunoprecipitation, and immunofluorescence. ..
- Mu J, Wang Y, Luo H, Leng M, Zhang J, Yang T, et al. A proteomic analysis of ataxia telangiectasia-mutated (ATM)/ATM-Rad3-related (ATR) substrates identifies the ubiquitin-proteasome system as a regulator for DNA damage checkpoints. J Biol Chem. 2007;282:17330-4 pubmed
- Zeng R, Spolski R, Leonard W. Measurement of interleukin-21. Curr Protoc Immunol. 2007;Chapter 6:Unit 6.30 pubmed publisher..Finally, a method to assess IL-21-induced tyrosine phosphorylation of Stat3 in splenic CD8(+) T cells using a flow cytometry-based analysis is described. ..
- Matsubayashi H, Sese S, Lee J, Shirakawa T, Iwatsubo T, Tomita T, et al. Biochemical characterization of the Drosophila wingless signaling pathway based on RNA interference. Mol Cell Biol. 2004;24:2012-24 pubmed
- Riento K, Totty N, Villalonga P, Garg R, Guasch R, Ridley A. RhoE function is regulated by ROCK I-mediated phosphorylation. EMBO J. 2005;24:1170-80 pubmed..This is the first demonstration of an endogenous Rho family member being phosphorylated in vivo and indicates that phosphorylation is an important mechanism to control the stability and function of this GTPase-deficient Rho protein...
- Wasco M, Pu R. Comparison of PAX-2, RCC antigen, and antiphosphorylated H2AX antibody (gamma-H2AX) in diagnosing metastatic renal cell carcinoma by fine-needle aspiration. Diagn Cytopathol. 2008;36:568-73 pubmed publisher..A combination of all 3 markers identified 87% of cases, and failure to stain for both PAX-2 and gamma-H2AX suggests against, but does not disprove, a diagnosis of RCC. ..
- Intine R, Tenenbaum S, Sakulich A, Keene J, Maraia R. Differential phosphorylation and subcellular localization of La RNPs associated with precursor tRNAs and translation-related mRNAs. Mol Cell. 2003;12:1301-7 pubmed
- Alberta J, Segal R. Generation and utilization of phosphorylation state-specific antibodies to investigate signaling pathways. Curr Protoc Neurosci. 2001;Chapter 3:Unit 3.14 pubmed publisher..Such antibodies can be used to address fundamental questions about signal transduction pathways during physiologic events that cannot be resolved by more conventional methodologies. ..
- Komatsu S, Ikebe M. ZIP kinase is responsible for the phosphorylation of myosin II and necessary for cell motility in mammalian fibroblasts. J Cell Biol. 2004;165:243-54 pubmed..These results suggest that ZIP kinase is critical for myosin phosphorylation and necessary for cell motile processes in mammalian fibroblasts. ..
- Saleh S, Albert A, Large W. Obligatory role for phosphatidylinositol 4,5-bisphosphate in activation of native TRPC1 store-operated channels in vascular myocytes. J Physiol. 2009;587:531-40 pubmed publisher..Moreover, the permissive role of PIP(2) in SOC activation requires PKC-dependent phosphorylation of TRPC1. ..
- Dorfleutner A, Cho Y, Vincent D, Cunnick J, Lin H, Weed S, et al. Phosphorylation of AFAP-110 affects podosome lifespan in A7r5 cells. J Cell Sci. 2008;121:2394-405 pubmed publisher..Therefore, we hypothesize that AFAP-110 phosphorylation and/or dephosphorylation is involved in the regulation of podosome stability and lifespan. ..
- Canesi L, Lorusso L, Ciacci C, Betti M, Gallo G. Effects of the brominated flame retardant tetrabromobisphenol-A (TBBPA) on cell signaling and function of Mytilus hemocytes: involvement of MAP kinases and protein kinase C. Aquat Toxicol. 2005;75:277-87 pubmed
- Rousseau S, Peggie M, Campbell D, Nebreda A, Cohen P. Nogo-B is a new physiological substrate for MAPKAP-K2. Biochem J. 2005;391:433-40 pubmed..The anisomycin-induced phosphorylation of Ser107 in HeLa cells can be prevented by 'knockdown' of MAPKAP-K2 using siRNA (small interfering RNA). Taken together, our results identify Nogo-B as a new physiological substrate of MAPKAP-K2. ..
- Ray H, Suau F, Vincent A, Dalla Venezia N. Cell cycle regulation of the BRCA1/acetyl-CoA-carboxylase complex. Biochem Biophys Res Commun. 2009;378:615-9 pubmed publisher..Taken together, our findings reveal a novel mechanism of regulation of ACCA distinct from the previously described phosphorylation of Ser79, and provide new insights into the control of lipogenesis through the cell cycle. ..
- Hutti J, Turk B, Asara J, Ma A, Cantley L, Abbott D. IkappaB kinase beta phosphorylates the K63 deubiquitinase A20 to cause feedback inhibition of the NF-kappaB pathway. Mol Cell Biol. 2007;27:7451-61 pubmed..Phosphorylation of A20 by IKKbeta thus represents part of a novel feedback loop that regulates the duration of NF-kappaB signaling following activation of innate immune signaling pathways. ..
- Oktay K, Oktay M. Immunohistochemical analysis of tyrosine phosphorylation and AP-1 transcription factors c-Jun, Jun D, and Fos family during early ovarian follicle development in the mouse. Appl Immunohistochem Mol Morphol. 2004;12:364-9 pubmed..Collectively, these data indicate that phosphotyrosine signaling and AP-1 transcription factors are intimately involved in early stages of ovarian follicle growth. ..
- Izawa I, Inagaki M. [Development of site- and phosphorylation state-specific antibodies]. Tanpakushitsu Kakusan Koso. 2005;50:1555-62 pubmed
- Swamy M, Schamel W. Purification of the T cell antigen receptor and analysis by blue-native PAGE. Methods Mol Biol. 2009;514:135-50 pubmed publisher..Using this powerful approach, the size, subunit composition and the relative abundance of the different TCR forms can be studied. We present here four methods to isolate the TCR in a native form and details to analyse it by BN-PAGE. ..
- Kim J, Kawasaki I, Shim Y. cdc-25.2, a C. elegans ortholog of cdc25, is required to promote oocyte maturation. J Cell Sci. 2010;123:993-1000 pubmed publisher..3. We propose that cdc-25 family members function as distinct but related cell cycle regulators to control diverse cell cycles in C. elegans germline development. ..
- Waraich R, Zaidi N, Moeschel K, Beck A, Weigert C, Voelter W, et al. Development and precise characterization of phospho-site-specific antibody of Ser(357) of IRS-1: elimination of cross reactivity with adjacent Ser(358). Biochem Biophys Res Commun. 2008;376:26-31 pubmed publisher..This antibody can be effectively used to further clarify the inhibitory role of Ser(357) in insulin signal transduction. ..
- Tootle T, Silver S, Davies E, Newman V, Latek R, Mills I, et al. The transcription factor Eyes absent is a protein tyrosine phosphatase. Nature. 2003;426:299-302 pubmed
- Toker A. PDK-1 and protein kinase C phosphorylation. Methods Mol Biol. 2003;233:171-89 pubmed
- Sefton B. Detection of phosphorylation by immunological techniques. Curr Protoc Protein Sci. 2001;Chapter 13:Unit13.4 pubmed publisher..This unit presents a protocol employing anti-phosphotyrosine antibodies with detection by either (125)I-labeled protein A or enhanced chemiluminescence (ECL). ..
- Schwertz H, Carter J, Abdudureheman M, Russ M, Buerke U, Schlitt A, et al. Myocardial ischemia/reperfusion causes VDAC phosphorylation which is reduced by cardioprotection with a p38 MAP kinase inhibitor. Proteomics. 2007;7:4579-88 pubmed..Thus, p38 MAP kinase inhibition with PD169316 is cardioprotective, reduces neutrophil activation, and controls protein expression and phosphorylation in MI and reperfusion. ..
- Galic S, Hauser C, Kahn B, Haj F, Neel B, Tonks N, et al. Coordinated regulation of insulin signaling by the protein tyrosine phosphatases PTP1B and TCPTP. Mol Cell Biol. 2005;25:819-29 pubmed..These results demonstrate that PTP1B and TCPTP are not redundant in insulin signaling and that they act to control both common as well as distinct insulin signaling pathways in the same cell. ..
- Williams C, Basu A, El Gharbawy A, Carrier L, Smith C, Rowan B. Identification of four novel phosphorylation sites in estrogen receptor alpha: impact on receptor-dependent gene expression and phosphorylation by protein kinase CK2. BMC Biochem. 2009;10:36 pubmed publisher..These novel ERalpha phosphorylation sites represent new means for modulation of ERalpha activity. S559 represents the first phosphorylation site identified in the extreme C-terminus (F domain) of a steroid receptor. ..
- Xu M, Ghosh I, Kochinyan S, Sun L. Intein-mediated peptide arrays for epitope mapping and kinase/phosphatase assays. Methods Mol Biol. 2007;381:313-38 pubmed..In this report, several experimental procedures including epitope mapping and the study of protein modifications were described. ..
- Goto H, Inagaki M. Production of a site- and phosphorylation state-specific antibody. Nat Protoc. 2007;2:2574-81 pubmed..This protocol is also applicable to the production of other types of antibodies, which specifically recognize the site-specific modification, such as acetylation, methylation and proteolysis. The protocol can be completed in 2-3 months. ..
- Litovchick L, Chestukhin A, DeCaprio J. Glycogen synthase kinase 3 phosphorylates RBL2/p130 during quiescence. Mol Cell Biol. 2004;24:8970-80 pubmed..Regulation of p130 by GSK3 provides a novel link between growth factor signaling and regulation of the cell cycle progression and exit. ..
- D Andrea M, Mei J, Tuman R, Galemmo R, Johnson D. Validation of in vivo pharmacodynamic activity of a novel PDGF receptor tyrosine kinase inhibitor using immunohistochemistry and quantitative image analysis. Mol Cancer Ther. 2005;4:1198-204 pubmed..In addition to this specific application of this in vivo validation approach to those targets that use PLCgamma as a downstream signaling partner, these methods may also benefit other drug discovery targets. ..
- He H, Tan M, Pamarthy D, Wang G, Ahmed K, Sun Y. CK2 phosphorylation of SAG at Thr10 regulates SAG stability, but not its E3 ligase activity. Mol Cell Biochem. 2007;295:179-88 pubmed..These studies suggested that CK2 might regulate SAG-SCF E3 ligase activity through modulating SAG's stability, rather than its enzymatic activity directly. ..
- Shi Y, Dodson G, Mukhopadhyay P, Shanware N, Trinh A, Tibbetts R. Identification of carboxyl-terminal MCM3 phosphorylation sites using polyreactive phosphospecific antibodies. J Biol Chem. 2007;282:9236-43 pubmed..Our findings also suggest that DSQ motifs are common phosphoacceptor motifs for ATM family kinases. ..
- Karwowska Desaulniers P, Ketko A, Kamath N, Pflum M. Histone deacetylase 1 phosphorylation at S421 and S423 is constitutive in vivo, but dispensable in vitro. Biochem Biophys Res Commun. 2007;361:349-55 pubmed..Taken together, the data are consistent with constitutive phosphorylation of HDAC1 at S421 and S423 in vivo, which is dispensable for activity in vitro. ..
- El Shemerly M, Hess D, Pyakurel A, Moselhy S, Ferrari S. ATR-dependent pathways control hEXO1 stability in response to stalled forks. Nucleic Acids Res. 2008;36:511-9 pubmed..The antibody may be a useful tool to monitor signal transduction events triggered by stalled DNA replication. ..
- Miyata Y, Nishida E. Evaluating CK2 activity with the antibody specific for the CK2-phosphorylated form of a kinase-targeting cochaperone Cdc37. Mol Cell Biochem. 2008;316:127-34 pubmed publisher..These results show that the anti-[pSer13]-Cdc37 antibody can be a promising new tool to evaluate in vivo CK2 activity. ..
- Wang L, Kobayashi T, Piao X, Shiono M, Takagi Y, Mineki R, et al. Serine 62 is a phosphorylation site in folliculin, the Birt-Hogg-Dubé gene product. FEBS Lett. 2010;584:39-43 pubmed publisher..A phospho-mimic mutation at Ser62 enhanced the formation of the FLCN-AMPK complex. These results suggest that function(s) of FLCN-AMPK-FNIP complex is regulated by Ser62 phosphorylation. ..
- Inoko A, Goto H. [Intermediate filaments: the mechanisms for the assembly and the physiological functions]. Tanpakushitsu Kakusan Koso. 2006;51:551-8 pubmed
- Malathi K, Kohyama S, Ho M, Soghoian D, Li X, Silane M, et al. Inositol 1,4,5-trisphosphate receptor (type 1) phosphorylation and modulation by Cdc2. J Cell Biochem. 2003;90:1186-96 pubmed..Taken together, these results indicate that IP3R1 may be a specific target for cdc2/CyB during cell cycle progression. ..