PhosphorylationSpecificityAntibody detects endogenous levelEndogenous levelsAntibodies were removedAFFINITY-CHROMATOGRAPHYEnrichmentPeptidesProteomic analysisSyntheticPeptideAssayPhosphoproteinsProteinIMACAbundanceMolecularHumanSignalCellSiteMethodSensitiveBandEndogenousPhosphorylation sitesAntibody detectsAntibodiesAffinity columnsPeptideAntigenAmino acidsSpectraLevelsCharacterizationProteinsPancreasFragmentationAnalysisSpecificHighlyWesternAbilityHeartEffectDetermine
Phosphorylation7
- Eighty-nine of these phosphoproteins were only detected in shortening flagella, whereas 29 were solely in flagella of steady growing cells, indicating dramatic changes of protein phosphorylation during flagellar shortening. (nih.gov)
- The experimentally validated phosphorylation sites as well as their catalytic kinases were extracted from public resources, and MS/MS phosphopeptides were manually curated from research articles. (nih.gov)
- Analyzing 300 µg of peptides per sample, we identified 22,712 phosphorylation sites, of which 19,075 were localized with high confidence and 1,203 are phosphorylated tyrosine residues, representing 6.3% of all detected phospho-sites. (scilifelab.se)
- The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Rb protein. (abcam.cn)
- DrJagannadham is the author of Detecting the site of phosphorylation in phospho peptides without loss of phosphate group using MALDI TOF mass spectrometry which was recently published in Analytical Chemistry Insights. (la-press.org)
- Fast Activated Cell-based ELISA (FACE™) Kits provide a simple, sensitive method for detecting protein phosphorylation directly in the cell, without making extracts or performing electrophoresis and membrane blotting. (activemotif.com)
- Altered phosphorylation of CREB phosphopeptide, a substrate of GSK3β, suggested that GSK3β may be the mediator of Cdk5 activity in kinesin-mediated axonal transport. (frontiersin.org)
Specificity1
- Specificity This antibody detects endogenous human, mouse, and rat GATA1 only when phosphorylated at serine 310. (qedbio.com)
Antibody detects endogenous level1
- This antibody detects endogenous level of AFX only when phosphorylated at serine 197. (prosci-inc.com)
Endogenous levels2
- Phospho-Chk2 (Thr383) Antibody detects endogenous levels of Chk2 only when phosphorylated at Threonine 383. (affbiotech.com)
- Phospho-STAT6 (Thr645) Antibody detects endogenous levels of STAT6 only when phosphorylated at Threonine 645. (affbiotech.cn)
Antibodies were removed1
- Non-phospho specific antibodies were removed by chromatogramphy using non-phosphopeptide. (prosci-inc.com)
AFFINITY-CHROMATOGRAPHY1
- Antibodies were purified by affinity-chromatography using epitope-specific phosphopeptide. (prosci-inc.com)
Enrichment6
- The use of 1,1,1,3,3,3-hexa-fluoroisopropanol is shown here to significantly improve Fe(III)-IMAC enrichment and subsequent detection of phosphopeptides by MALDI-MS. (reading.ac.uk)
- The selectivity of the amphiphilic nanoassemblies were changed by loading metal ions for the enrichment of phosphopeptides and by incorporating hydrazide functional groups for the enrichment of glycopeptides. (umass.edu)
- In addition to developing new nanoassemblies for the enrichment of phosphopeptides and glycopeptides, we also explored how supramolecular systems could be tuned to enhance extraction selectivity and efficiency via structural variations to the amphiphilic polymers. (umass.edu)
- The utility of these materials for the enrichment of phosphopeptides and glycopeptides from complexed samples is also demonstrated. (umass.edu)
- Phosphopeptide enrichment and Tandem mass tags labeling For PS1, SGK3 knock-out HEK 293 (SGK3 knock-out) cells were generated by the Crispr/Cas9 methodology as described earlier. (baxkyardgardener.com)
- Phosphopeptide enrichment For phosphopeptide enrichment, titanium oxide (TiO2) beads (Titansphere Phos-TiO2 Bulk 10?m #5010C21315, GL Sciences, Japan) were used [34,35] and prepared by washing with 100% acetonitrile. (baxkyardgardener.com)
Peptides4
- IMAC can be used to selectively enrich phosphopeptides from complex peptide mixtures, but co-retention of acidic peptides together with the failure to retain some phosphopeptides restricts the general utility of the method. (reading.ac.uk)
- Here we demonstrate that, as expected, peptides with a more acidic amino acid content are generally more efficiently purified and detected by MALDI-MS after Fe(III)-IMAC than those with a more basic content. (reading.ac.uk)
- However, detecting PTMs on proteins and peptides by MS is challenging because of their low abundance and heterogeneity. (umass.edu)
- HiRIEF fractions with the most acidic isoelectric points are enriched in multiply phosphorylated peptides, which represent 18% of all the phospho-peptides detected in the pH range 2.5-3.7. (scilifelab.se)
Proteomic analysis1
Synthetic2
- In this study Fe(III)-IMAC was qualitatively and quantitatively assessed using a panel of phosphopeptides, both synthetic and derived from proteolysis of known phosphoproteins, to identify the causes of success and failure in the application of this technique. (reading.ac.uk)
- Antibodies were produced by immunizing rabbits with synthetic phosphopeptide and KLH conjugates. (sabbiotech.cn)
Peptide1
- A fluorescent peptide is used in biological experiments to detect a reaction or change. (nridigital.com)
Assay2
- The assay detected signal across at least 3 orders of magnitude of lysate input with a linear range spanning 1-2 orders and a low femtomole limit of detection for EGFR phosphotyrosine. (biomedcentral.com)
- FACE is a highly sensitive high-throughput assay designed for detecting activated proteins within mammalian cells. (activemotif.com)
Phosphoproteins2
- A total of 224 phosphoproteins derived from 1296 spectral counts of phosphopeptides were identified. (nih.gov)
- This analysis identified 9,643 phosphopeptides from 2,413 proteins, making it the "largest group of EV phosphoproteins [identified] to date," the authors wrote. (genomeweb.com)
Protein5
- A single protein band was detected. (antikoerper-online.de)
- A single protein band was detected from all three lysates. (antikoerper-online.de)
- 4) Positive and Negative Controls: Mouse Anti-GAPDH antibody (included) should be used to detect the internal positive controls for normalization of OD values of the target protein. (antikoerper-online.de)
- This antibody detects endogenous protein at a molecular weight of 90 kDa. (thermofisher.com)
- 3] Because the vast majority of mutations reside in and around the broad intramolecular interaction surface between the N-SH2 and PTP domains of the PTPN11 protein, they have been suggested to affect the intramolecular N-SH2/PTP binding in the absence of a phosphopeptide, leading to excessive phosphatase activities. (medscape.com)
IMAC2
- The phosphopeptides were enriched by a combination of IMAC and titanium dioxide chromatography with a strategy of sequential elution from IMAC (SIMAC) and analyzed by tandem mass spectrometry. (nih.gov)
- Modulating the loading buffer used for Fe(III)-IMAC significantly affects phosphopeptide binding and suggests that conformational factors that lead to steric hindrance and reduced accessibility to the phosphate are important. (reading.ac.uk)
Abundance3
- When they targeted 101 phosphopeptides that they were interested in -- some of which were decent abundance, some of them were exceptionally low abundance (DDA and DIA didn't even pick them up! (blogspot.com)
- However, the differential abundance of multiple CREB1 phosphopeptides suggested an increase in CREB1 activity in the higher producing cell line, which was confirmed by increased association of the CMV promotor with CREB1 in the high producer cell line. (inpst.net)
- One of the challenges of discovering and developing circulating biomarkers is the complexity of plasma, which makes reproducibly detecting disease-linked markers, which are often low abundance, a difficult analytical problem. (genomeweb.com)
Molecular2
- Detects a band of approximately 120 kDa (predicted molecular weight: 106 kDa). (abcam.cn)
- Three radioactive components of apparent molecular masses of 100 kD, 76 kD, and 26 kD were detected. (uni-bielefeld.de)
Human3
- Detects human EGF R phosphorylated at Y1068. (rndsystems.com)
- The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human Syk that contains tyrosine 323 (tyrosine 317 in the mouse sequence). (thermofisher.cn)
- Amplification of the ErbB-2 gene and overexpression of its product has been detected in almost 40% of primary human breast tumors, which correlates with poor prognosis in node positive breast cancer. (activemotif.com)
Signal2
- Signal is detected either by chemiluminescence (far-Western and rosette) or by PCR (OTM). (biomedcentral.com)
- The signal was detected using a Li-Cor Biosciences Odyssey System and quantified in Image Studio Lite (Li-Cor) or using the ECL Western Blotting Detection Kit (Amersham) on Amersham Hyperfilm ECL films (Amersham). (baxkyardgardener.com)
Cell3
- The ETK (Phospho-Tyr566) Colorimetric Cell-Based ELISA Kit can detect Phospho-ETK expression in as little as 5,000 HeLa cells. (antikoerper-online.de)
- They take a complex phosphoproteomic model (MCF-7 breast cancer cell line) and look at the phosphoproteome using normal (difficult and described above) shotgun DDA techniques, then DIA (like that SWATH thing), and finally with Parallel Reaction Monitoring (PRM) for targeted phosphopeptides of interest. (blogspot.com)
- Mimotopes offers phosphopeptide synthesis for cell research, as well as phosphopeptides to be used as antigens in the production of anti-phosphopeptide antibodies. (nridigital.com)
Site1
- On hand from the data it can be hypothesised that the 76 kD phosphopeptide may be the phosphointermediate of the H+-ATPase that has been demonstrated in the PM of T. viridis (Popova and Balnokin, 1992), while the 26 kD polypeptide may participate in the catalytic cycles of the ATPases, being a primary site of phosphate binding. (uni-bielefeld.de)
Method1
- If your goal is to get a method that gets more phosphopeptides than the last paper, more power to you. (blogspot.com)
Sensitive1
- The 100 kD phosphopeptide was sensitive to ADP, which caused rapid discharge of the phosphopeptide. (uni-bielefeld.de)
Band1
- A band of ~43kDa is detected. (qedbio.com)
Endogenous8
- The phospho-glycogen synthase antibody (Tyr44) detects endogenous levels of glycogen synthase only when it is phosphorylated at Tyr44. (nate-trust.org)
- SMC1 antibody (Ser966) detects endogenous levels of SMC1 only when phosphorylated at serine 966. (mc1-r.com)
- Phospho-Akt (T308) Polyclonal Antibody detects endogenous levels of Akt protein only when phosphorylated at T308. (bionewway.com)
- Bamacan (Phospho-Ser1067) Antibody detects endogenous levels of Bamacan only when phosphorylated at Ser1067. (researchgrade.com)
- Phospho-MAP3K8/COT (Thr290) Antibody detects endogenous levels of MAP3K8/COT only when phosphorylated at Threonine 290. (affbiotech.com)
- Phospho-Dynamin 1 (Ser774) Antibody detects endogenous levels of Dynamin 1 only when phosphorylated at Serine 774. (affbiotech.cn)
- Phospho-NR3C1 (Ser203) Antibody detects endogenous levels of NR3C1 only when phosphorylated at Sersine 203. (affbiotech.com)
- Phospho-Trk B (Y706/Y707) Polyclonal Antibody detects endogenous levels of Trk B protein only when phosphorylated at Y706/Y707. (immunoway.com)
Phosphorylation sites3
- Demonstrating this, the study, the first to be published using TMTcalibrator™, reports the detection of 47 tau phosphopeptides covering 31 clinically-relevant phosphorylation sites. (proteomics.com)
- Daporinad molecular weight Casein bands Selleck PKC412 were digested with trypsin and the resulting peptides were analyzed by nLC MS/MS. Eight out of nine predicted tryptic phosphopeptides corresponding to 18 different phosphorylation sites were detected in alpha(s1)-, a(s2)-, and beta-casein. (pka-signal.com)
- 7. Simultaneous quantification of protein phosphorylation sites using liquid chromatography-tandem mass spectrometry-based targeted proteomics: a linear algebra approach for isobaric phosphopeptides. (nih.gov)
Antibody detects3
- The antibody detects phosphorylated Asc (Tyr-144) in the inflammasome. (ecmbio.com)
- In addition, the antibody detects a 20 kDa Asc protein in western blots of J774A.1 mouse macrophages treated with pervanadate. (ecmbio.com)
- The antibody detects a 116 kDa* band corresponding to PYK2 on SDS-PAGE immunoblots of human A431, Jurkat and K562 cells treated with pervanadate, but is not observed in control, untreated cells. (ecmbio.com)
Antibodies4
- Phosphopeptide antibodies and phosphomimetic mutants of these nuclear receptors were utilized to examine whether phosphorylation becomes a regulatory factor determining the glucose responsive SULT1E1 gene activation. (nih.gov)
- We began assembling other reports of deleterious interactions between biomolecules and high performance liquid chromatography (HPLC) systems, cataloging problems with phosphopeptides (1,2), phosphorylated glycans (3), monoclonal antibodies (mAbs) (4), and pharmaceutical compounds (5). (chromatographyonline.com)
- With PTMScan, instead of an antibody specific to a single phosphopeptide sequence, a peptide-library-based approach is used for antibody development, which generates antibodies specific to just the phosphorylated amino acid residue (or any other PTM residue). (cellsignal.com)
- We provide tools for path-breaking research by developing antibodies that detect a comprehensive library of novel and established targets. (abcepta.com)
Affinity columns2
- The antibody is derived from rabbit serum purified by affinity purification by sequential chromatography on phosphopeptide and non-phosphopeptide affinity columns. (nate-trust.org)
- Prepared from pooled rabbit serum by affinity purification via sequential chromatography on phospho and non-phosphopeptide affinity columns. (phosphosolutions.com)
Peptide3
- Peptide sequence information can be directly obtained from phosphopeptides MS2 spectra. (bruker.com)
- Send us an aliquot of your antibody and the amino acid sequence of the binding partner and we will perform peptide synthesis, detect epitope with your antibody and data interpretation, and provide a complete documentation of the results. (abnova.com)
- In the example below, a single phosphopeptide is used as the antigen, and an antibody is developed that is specific only to a peptide sequence with the defined phosphotyrosine residue. (cellsignal.com)
Antigen3
- It detects mouse and rat Serotonin Transporter (Thr276) and is antigen affinity purified from pooled serum. (phosphosolutions.com)
- Phosphospecificity is shown in the second lane (+) where Immunolabeling is blocked by preadsorption of the phosphopeptide used as antigen, but not by the corresponding non-phosphopeptide (not shown). (phosphosolutions.com)
- Immunolabeling is blocked by preadsorption with the phosphopeptide used as antigen, but not by the corresponding non-phosphopeptide. (phosphosolutions.com)
Amino acids1
- Clone M269 was generated from a phosphopeptide containing amino acids surrounding tyrosine 402 in human PYK2. (ecmbio.com)
Spectra4
- 1. Colander: a probability-based support vector machine algorithm for automatic screening for CID spectra of phosphopeptides prior to database search. (nih.gov)
- 2. Automatic validation of phosphopeptide identifications from tandem mass spectra. (nih.gov)
- 12. A hierarchical MS2/MS3 database search algorithm for automated analysis of phosphopeptide tandem mass spectra. (nih.gov)
- 15. Effect of high-accuracy precursor masses on phosphopeptide identification from MS3 spectra. (nih.gov)
Levels1
- Of the 28 HAfTs that were initially detected, the identity, presence, and levels of 17 were confirmed by Sanger sequencing or polymerase chain reaction. (nih.gov)
Characterization1
- 2017. Characterization of the APLF FHA-XRCC1 phosphopeptide interaction and its structural and functional implications. (nih.gov)
Proteins3
- We detected the expected phosphorylation of MAP kinases, translational regulatory proteins, and subunits of APC/C that validate the accuracy of our measurements. (xenbase.org)
- Proteins that were not detected have had their abundance set to zero. (xenbase.org)
- The ability to accurately detect, profile and characterize post translational modifications (PTMs) on proteins in a sample is essential to the understanding of biological systems. (bruker.com)
Pancreas1
- Isoform TrkB-T1 is mainly expressed in the brain but also detected in other tissues including pancreas, kidney and heart. (immunoway.com)
Fragmentation1
- 16. Automated phosphopeptide identification using multiple MS/MS fragmentation modes. (nih.gov)
Analysis2
- Well established methods involving the analysis of 32 P-labeled phosphoproteins by Edman degradation and two-dimensional phosphopeptide mapping have proven to be powerful but not without limitations. (biomedcentral.com)
- 18. Automated platform of μLC-MS/MS using SAX trap column for highly efficient phosphopeptide analysis. (nih.gov)
Specific1
- No metal binding or enzymatic activity can be detected for Memo, but Memo does bind directly to a specific ErbB2-derived phosphopeptide encompassing Tyr-1227 using its vestigial enzymatic active site. (crcm-marseille.fr)
Highly1
- Unexpectedly, we find that β-arrestin binding to phosphorylated receptor, with a phosphorylation barcode identical to the isolated phosphopeptide, is highly inefficient and that agonist-promoted receptor activation is required for β-arrestin activation, consistent with the release of a sequestered receptor C tail. (nih.gov)
Western1
- Expression of molecules such as connective tissue growth factor (CTGF), SMAD family members 3 (Smad3) and 7 (Smad7), and small heterodimer partner were detected by RT-PCR and Western blotting as part of the application. (bvsalud.org)
Ability1
- These include the limited ability to detect early stage PD prior to the onset of motor signs, inadequate measures of processes or pathways related to disease pathogenesis, and the lack of biomarkers that define disease progression. (nih.gov)
Heart1
- In the heart, detected in left and right atria, left and right ventricles, aorta, apex, intraventricular septum, and atrioventricular node as well as whole adult and fetal heart (PubMed:10902803). (affbiotech.com)
Effect1
- Makes antineoplastic AZD2171 in several tumors, including normal lung, hepatocellular Rem cases cancer and prostate cancer as well as in all the F, The antitumor effect was detected associated with a strong inhibition of VEGF signaling and angiogenesis. (survivinpathway.com)
Determine1
- The antibody can be used in immunoassays to detect ATR in a sample, to determine the inhibitory activity of ATR agents, and to measure the effectiveness of an ATR modulator agent or ATR inhibitor as therapeutics. (nih.gov)