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Phosphotyrosine

Phosphotyrosine is a chemical modification of the amino acid tyrosine, where a phosphate group is added to the hydroxyl group of the tyrosine residue.
This posttranslational modification plays a crucial role in cellular signaling pathways, regulating protein-protein interactions, enzyme activity, and subcellular localization.
Phosphotyrosine-containing proteins are involved in a wide range of biological processes, including cell growth, differentiation, metabolism, and immune response.
Researchers studying phosphotyrosine-mediated signaling can leverage PubCompare.ai's AI-driven tools to streamline their experiments, locate the best protocols from literature, pre-prints, and patents, and optimize their research.
With PubCompare.ai's intuitive interface, scientists can take their phosphotyrosine research to the next leval.

Most cited protocols related to «Phosphotyrosine»

1
Immunoblotting Analysis of Signaling Proteins
Cited 8 times
For immunoblotting, 100 μg of cell lysates were resolved in an SDS-PAGE and Western immunoblotting with antibody as indicated in the figure legend. Src monoclonal antibodies GD11 and 2-17 were generously provided by Dr. Sarah J. Parsons. Antibodies against actin, amino-terminal FAK (A17), and phosphotyrosine (PY20) were from Santa Cruz Biotechnology (Santa Cruz, California); and the antibody against phosphorylated Tyr- 397 of FAK was from Upstate Biotechnology, Inc. (Lack Placid, NY). The E10 monoclonal antibody recognized phosphorylated Thr-202 and Tyr-204 of ERK1/2 was purchased from New England Biolabs, Inc. (Beverly, MA, USA). The rabbit polyclonal antibodies recognized unphosphorylated ERK2 were purchased from Santa Cruz. Western immunoblotting was performed with antibody of interest and detected by enhanced chemiluminescence (Amersham Biosciences).
Lin T.Y., Fan C.W., Maa M.C, & Leu T.H. (2015). Lipopolysaccharide-promoted proliferation of Caco-2 cells is mediated by c-Src induction and ERK activation. BioMedicine, 5(1), 5.
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Publication 2015
Actins Antibodies Cells Chemiluminescence Immunoglobulins Mitogen-Activated Protein Kinase 3 Mitogen Activated Protein Kinase 1 Monoclonal Antibodies Phosphotyrosine Rabbits SDS-PAGE
2
Immunoblot Analysis of Protein Signaling
Cited 8 times
Cells were lysed with lysis buffer (50 mM Tris-HCl, pH 8.0; 150 mM NaCl, 0.1% NP-40, 0.5% CHAPS) containing 0.1% protease inhibitor cocktail III (Calbiochem, San Diego, CA). The lysates were electrophoretically separated, blotted onto a nitrocellulose membrane and blocked with 4% BlockAce solution (Dainippon Pharmaceutical, Osaka, Japan) for 1 h. The blots were then incubated with antibodies against the following proteins: BIG3 (ref. 21 (link)) (1:200); PHB2 (1:500), NcoR (1:500) and ERα (phospho Y537; 1:500; Abcam, Cambridge, UK); SRC-1 (128E7; 1:500), Shc (1:500), α/β-tubulin (1:1,000), Akt (1:1,000), phospho-Akt (S473; 587F11; 1:1,000), p44/42 MAPK (1:500), phospho-p44/42 MAPK (T202/Y204; 1:500) and phospho-ERα (S104/S106; 1:500; Cell Signaling Technology, Danvers, MA); HDAC1 (H-11; 1:500), IGF-1Rβ (1:500), PI3-kinase p85α (U13; 1:500), phospho-ERα (S118; 1:500), phospho-ERα (S167; 1:500) and laminin B1 (1:100; Santa Cruz Biotechnology, Santa Cruz, CA); phospho-ERα (S305; 1:500; Millipore, Billerica, MA); phosphotyrosine (1:500; Zymed, San Francisco, CA); β-actin (AC-15; 1:5,000) and FLAG-tag M2 (1:5,000; Sigma, St Louis, MO); and HA-tag (1:3,000; Roche, Mannheim, Germany). After incubation with an horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotechnology, dilution 1:5,000) or monoclonal anti-rabbit immunoglobulins-peroxidase antibody (RG-16, Sigma, dilition 1:5,000) for 1 h, the blots were developed with an enhanced chemiluminescence system (GE Healthcare, Buckinghamshire, UK) and were scanned using an Image Reader LAS-3000 mini (Fujifilm, Tokyo, Japan). All experiments were performed more than three times in triplicate. Finally, the phosphorylation levels of IGF-1Rβ, Shc, PI3K, Akt, p42/44 MAPK and ERα were assessed through densitometric analysis of immunoblot results using an Image Reader LAS-3000 mini51 . Full-length images of immunoblots are shown in Supplementary Fig. S9.
Yoshimaru T., Komatsu M., Matsuo T., Chen Y.A., Murakami Y., Mizuguchi K., Mizohata E., Inoue T., Akiyama M., Yamaguchi R., Imoto S., Miyano S., Miyoshi Y., Sasa M., Nakamura Y, & Katagiri T. (2013). Targeting BIG3–PHB2 interaction to overcome tamoxifen resistance in breast cancer cells. Nature Communications, 4, 2443.
Publication 2013
1-Phosphatidylinositol 3-Kinase 3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate Actins Antibodies Antibodies, Anti-Idiotypic Buffers Cells Chemiluminescence Densitometry Horseradish Peroxidase Immunoblotting Immunoglobulins Laminin Mitogen-Activated Protein Kinase 3 Mitogen Activated Protein Kinase 1 Monoclonal Antibodies NCOA1 protein, human NCOR1 protein, human Nitrocellulose Nonidet P-40 Peroxidase Pharmaceutical Preparations PHB2 protein, human Phosphatidylinositol 3-Kinases Phosphorylation Phosphotyrosine pitrilysin Proteins Rabbits Sodium Chloride Technique, Dilution Tissue, Membrane Tromethamine Tubulin
3
Western Blot Analysis of Protein Interactions
Cited 8 times
Cell extracts or immunoprecipitated proteins were diluted in SDS sample buffer and boiled for 5 min. Proteins were separated on 10% SDS-PAGE gels and transferred to Hybond-P membranes (GE Healthcare) as previously described (Sette et al., 2002 (link)). The following primary antibodies (1:1,000 dilution) were used (overnight at 4°C): rabbit anti-Sam68, rabbit anti-Erk2, rabbit anti-Fyn, and mouse anti-phosphotyrosine (PY20; Santa Cruz Biotechnology, Inc.); mouse anti-ASF/SF2 (US Biological); mouse anti–hnRNP A1, mouse anti-tubulin, and rabbit anti-actin (Sigma-Aldrich); mouse anti–hnRNP H/F (Abcam); rabbit anti–phospho-ERKs (Cell Signaling); rabbit anti-GFP (Roche); rabbit anti-phosphoserine and anti-phosphothreonine (Stressgen); and rabbit anti–Bcl-x (BD Biosciences). Secondary anti-mouse or anti-rabbit IgGs conjugated to horseradish peroxidase (GE Healthcare) were incubated with the membranes for 1 h at room temperature at a 1:10,000 dilution in PBS containing 0.1% Tween 20. Immunostained bands were detected by chemiluminescent method (Santa Cruz Biotechnology, Inc.).
Paronetto M.P., Achsel T., Massiello A., Chalfant C.E, & Sette C. (2007). The RNA-binding protein Sam68 modulates the alternative splicing of Bcl-x. The Journal of Cell Biology, 176(7), 929-939.
Publication 2007
Actins anti-IgG Antibodies bcl-X Protein Biopharmaceuticals Buffers Cell Extracts EPHB2 protein, human Gels Heterogeneous-Nuclear Ribonucleoprotein Group F Heterogeneous Nuclear Ribonucleoprotein A1 Horseradish Peroxidase Mitogen Activated Protein Kinase 1 Mus Phosphoserine Phosphothreonine Phosphotyrosine Proteins Rabbits SDS-PAGE Technique, Dilution Tissue, Membrane Tubulin Tween 20
4
Tyrosine Kinase Receptor Signaling Assay
Cited 7 times
697 B-ALL cells
and Molm-14 AML cells were cultured in the presence of 11 or vehicle-only for 1.0 h. Pervanadate solution was prepared fresh
by combining 20 mM sodium orthovanadate in 0.9× PBS in a 1:1
ratio with 0.3% (w/w) hydrogen peroxide in PBS for 15–20 min
at room temperature. Cultures were treated with 120 μM pervanadate
for 3 min prior to collection, and cell lysates were prepared in 50
mM HEPES (pH 7.5), 150 mM NaCl, 10 mM EDTA, 10% glycerol, and 1% Triton
X-100, supplemented with protease inhibitors (Roche Molecular Biochemicals,
no. 11836153001). Mer and Flt3 proteins were immunoprecipitated with
anti-Mer (R&D Systems, no. MAB8912) or anti-Flt3 (Santa Cruz Biotechnology
no. sc-480) antibody and Protein G agarose beads (InVitrogen). Phospho-proteins
were detected by Western blot using an antiphospho-Mer antibody raised
against a peptide derived from the triphosphorylated activation loop
of Mer8 (link) (Phopshosolutions, Inc.) or an
antibody specific for phosphorylated Flt3 (Cell Signaling Technology,
no. 3461). Nitrocellulose membranes were stripped and total proteins
were detected using a second anti-Mer antibody (Epitomics Inc., no.
1633-1) or anti-Flt3 antibody (Santa Cruz Biotechnology no. sc-480).
Relative phosphorylated and total protein levels were determined by
densitometry using ImageJ, and IC50 values were calculated
by nonlinear regression.
32D Cells expressing a chimeric EGFR-Mer,
EGFR-Axl, or EGFR-Tyro3 receptor were cultured in the presence of 11 or vehicle-only for 1.0 h before stimulation with 100 ng/mL
EGF (BD Biosciences no. 354010) for 15 min. Cells were centrifuged
at 1000g for 5 min and washed with 1× PBS. Cell
lysates were prepared in 20 mM HEPES (pH 7.5), 50 mM NaF, 500 mM NaCl,
5.0 mM EDTA, 10% glycerol, and 1% Triton X-100, supplemented with
protease inhibitors (10 μg/mL leupeptin, 10 μg/mL phenylmethylsulfonyl
fluoride, and 20 μg/mL aprotinin) and phosphatase inhibitors
(50 mM NaF and 1.0 mM sodium orthovanadate). Mer protein was immunoprecipitated
using a custom polyclonal rabbit antisera raised against a peptide
derived from the C-terminal catalytic domain of Mer and Protein A
agarose beads (Santa Cruz Biotechnology). Axl and Tyro3 proteins were
immunoprecipitated using an antibody directed against a FLAG epitope
engineered into the chimeric proteins (Sigma-Aldrich, no. F1804).
Phosphotyrosine-containing proteins were detected by Western blot
with a monoclonal HRP-conjugated antiphosphotyrosine antibody (Santa
Cruz Biotechnology, no. sc-508). Antibodies were stripped from membranes,
and total proteins were detected with the same antibodies used for
immunoprecipitation.
Zhang W., DeRyckere D., Hunter D., Liu J., Stashko M.A., Minson K.A., Cummings C.T., Lee M., Glaros T.G., Newton D.L., Sather S., Zhang D., Kireev D., Janzen W.P., Earp H.S., Graham D.K., Frye S.V, & Wang X. (2014). UNC2025, a Potent and Orally Bioavailable MER/FLT3 Dual Inhibitor. Journal of Medicinal Chemistry, 57(16), 7031-7041.
Publication 2014
Antibodies Antibodies, Anti-Idiotypic Aprotinin Catalytic Domain Cells Chimera Edetic Acid EGFR protein, human FLT3 protein, human G-substrate Glycerin HEPES Immune Sera Immunoglobulins inhibitors Intestinal Atresia, Multiple leupeptin Monoclonal Antibodies Nitrocellulose Orthovanadate Peptides Peroxide, Hydrogen pervanadate Phosphoric Monoester Hydrolases Phosphotyrosine Protease Inhibitors Proteins Rabbits Sepharose Sodium Sodium-20 Sodium Chloride Tissue, Membrane Triton X-100 Western Blotting
5
Imaging of Actin Cytoskeleton Dynamics
Cited 7 times
The acetoxymethyl-ester of SNARF-5, nigericin, EGF, TMR-dextran (Mw 10,000), transferrin-Alexa 546, and rhodamine-phalloidin were from Invitrogen. HOE-694 was a gift from Dr. H.-J. Lang (Aventis Pharma, Frankfurt am Main, Germany). Lipofectamine LTX was from Invitrogen, Fugene6 from Roche, LY294002 from Enzo Life Sciences, Inc., latrunculin B from EMD, G-Sepharose beads from GE Healthcare, and rabbit skeletal muscle rhodamine-actin from Cytoskeleton, Inc. Mouse monoclonal anti-phosphotyrosine (4G10) and anti-GAPDH antibodies were from Millipore, rabbit monoclonal anti-phospho-Akt (Ser 473) antibody was from Cell Signaling Technology, mouse monoclonal anti-Rac1 and anti-Cdc42 were from BD, and rabbit polyclonal anti-cofilin and anti–phospho-cofilin were from Abcam. All other chemicals were from Sigma-Aldrich.
Isotonic Na+-rich buffer contained 140 mM NaCl, 3 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 5 mM glucose ,and 20 mM Hepes, pH 7.4. In NMG+-rich buffer NaCl and KCl were replaced by 143 mM NMG-chloride, and in K+-rich buffer NaCl was replaced by 100 mM K-glutamate and 43 mM KCl.
Koivusalo M., Welch C., Hayashi H., Scott C.C., Kim M., Alexander T., Touret N., Hahn K.M, & Grinstein S. (2010). Amiloride inhibits macropinocytosis by lowering submembranous pH and preventing Rac1 and Cdc42 signaling. The Journal of Cell Biology, 188(4), 547-563.
Publication 2010
Actin Depolymerizing Factors Anti-Antibodies Buffers CDC42 protein, human Chlorides Dextran Esters GAPDH protein, human Glucose Glutamates HEPES HOE 694 Immunoglobulins latrunculin B Lipofectamine LY 294002 Magnesium Chloride Microfilaments Mus Nigericin Phosphotyrosine Rabbits Rhodamine rhodamine-phalloidin Sepharose Skeletal Muscles Sodium Chloride Transferrin

Most recents protocols related to «Phosphotyrosine»

1
Quantitative BTK Phosphorylation Inhibition Assay
Not available on PMC !

Example 156

Human heparinized venous blood was purchased from Bioreclamation, Inc. or SeraCare Life Sciences and shipped overnight. Whole blood was aliquoted into 96-well plate and “spiked” with serial dilutions of test compound in DMSO or with DMSO without drug. The final concentration of DMSO in all wells was 0.1%. The plate was incubated at 37° C. for 30 min. Lysis buffer containing protease and phosphatase inhibitors was added to the drug-containing samples and one of the DMSO-only samples (+PPi, high control), while lysis buffer containing protease inhibitors was added to the other DMSO-only samples (−PPi, low control). All of the lysed whole blood samples were subjected to the total BTK capture and phosphotyrosine detection method described in US20160311802, incorporated herein by reference. ECL values were graphed in Prism and a best-fit curve with restrictions on the maximum and minimum defined by the +PPi high and −PPi low controls was used to estimate the test compound concentration that results in 50% inhibition of ECL signal by interpolation.

Table 2 shows the activity of selected compounds of this invention in the pBTK assay, wherein each compound number corresponds to the compound numbering set forth in Examples 1-154 described herein. “†” represents an IC50 of equal to or less than 10,000 nM but greater than 500 nM, “††” represents an IC50 of equal to or less than 500 nM but greater than 100 nM; and “†††” represents an IC50 of equal to or less than 100 nM.

TABLE 2
IC50 (nM)Compound No.
†††2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14a, 15, 16, 17, 18, 19, 20, 21, 22, 23,
24a, 25, 26a, 26b, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36a, 37, 39, 40, 41, 42a,
44, 56, 58, 60, 62, 63, 64, 65, 67, 68, 71, 72, 73, 75, 76, 77, 78, 79, 80, 81,
82, 83, 84, 86, 87, 88, 89a, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101,
104, 105, 106, 107, 110, 111, 112, 113, 114, 119a, 120, 121, 122, 123, 124,
125, 126, 152,
††38, 42b, 45, 46, 47, 48, 49, 51, 52, 53, 57, 59, 61, 66, 69, 70, 74, 102, 103,
115, 117, 118a, 118b, 127, 128, 129, 130, 131, 132
†14b, 24b, 36b, 43, 50, 85, 89b, 108, 109, 116, 133, 134, 135, 136, 137, 138,
139, 140, 141, 153,

US11858926B2. Inhibiting agents for bruton's tyrosine kinase (2024-01-02). BIOGEN MA INC. [US]. Inventors: Brian T. Hopkins [US], Bin Ma [US], Robin Prince [US], Isaac Marx [US], Joseph P. Lyssikatos [US], Fengmei Zheng [US], Matthew Peterson [US], Daniel B. Patience [US].
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Patent 2024
Biological Assay BLOOD Buffers Homo sapiens inhibitors Peptide Hydrolases Pharmaceutical Preparations Phosphoric Monoester Hydrolases Phosphotyrosine prisma Protease Inhibitors Psychological Inhibition Sulfoxide, Dimethyl Technique, Dilution Veins
2
Structural Comparison of Mammalian Numb Isoforms
Not available on PMC !

Example 37

Structural comparison between mouse Numblike and its mammalian Numb homologues and construction of integrase-deficient, transgene expressing lentivectors.

FIG. 20A illustrates that Numblike shows greater than 70% sequence identity in its amino terminal half to the shortest Numb homologue, but less than 50% identity in its cytoplasmic half where a unique 15 amino acid polyglutamine domain (purple) is found. The longest Numb isoform contains an 11 amino acid insert (white) within its phosphotyrosine binding (PTB) domain (black), as well as a 49 amino acid insert (gray) adjacent to a proline rich region (PRR). Two intermediate sized isoforms contain either the PTB or PRR inserts, but not both. The shortest Numb isoform lacks both inserts. FIG. 20B illustrates the HIV-EGFP Numblike and HIV-EGFP-NumbPTB+/PRR+vectors constructed from the two-gene HIV-EGFP-HSA vector (Reiser et al., 2000) by cloning the transgene cDNAs into nef coding region previously occupied by the mouse HSA cDNA. Abbreviations: Rev-response element (RRE), slice donor site (SD), splice acceptor site (SA).

US11859168B2. Electroporation, developmentally-activated cells, pluripotent-like cells, cell reprogramming and regenerative medicine (2024-01-02). None [US]. Inventors: Christopher B. Reid [US], Melissa Braga [US], Lilian N. Santamaria [US], Ashley N. Wickrema [US], Marinne D. Wickrema [US], Anthony Hao Dinh [US], Yaman Eksioglu [US], Mat Hoang Ho [US].
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Patent 2024
Amino Acids Cells Cloning Vectors Cytoplasm DNA, Complementary Electroporation Genetic Vectors Integrase Mammals Mice, Laboratory Phosphotyrosine polyglutamine Proline Protein Isoforms Response Elements Splice Acceptor Site Tissue Donors Transgenes
3
Recombinant Antibody and Protein Purification
HER2-specific antibodies Trastuzumab, 39 S, BS4 as well as TfR-specific antibodies 226, 289 and 292 were transiently expressed by co-transfection of the heavy and light chain expressing vectors into CHO cells using a polyethylenimine-based method46 (link). Supernatants were purified by affinity chromatography using a Protein A MabSelectSure column (GE Healthcare) and molecules were analysed for integrity by SDS-PAGE and SEC-HPLC using a TSKgel G3000SWxl column (Tosoh Bioscience). Wheat-germ-agglutinin (L9640) and biotin-transferrin (T3915) were from Merck. Antibodies/ligands were labelled with DyLight-650 or 488 NHS Ester (Thermo Fisher Scientific 62265 and 46402) according to manufacturers’ instructions. FabFluor Red Antibody Labelling Reagent (4722) was from Sartorius. Transferrin-AlexaFluor546 (T23364) and EGF-Alexa647-complex (E35351), Dextran (70 kDa)-Tetramethylrhodamine (D1818) and Dextran (70 kDa)-Fluorescein (D1822) were from Thermo Fisher Scientific. Antibodies recognising the HER2 cytoplasmic domain (2242), pHER2 (2241), HER3 (12708), the HA-tag (2367) and EGFR (4267) were from Cell Signaling Technology. An antibody recognising the HER2 ectodomain (AF1129) was from R&D Systems. Antibodies recognising VAV1 (ab245440), VAV2 (ab52640), VAV3 (ab52938), GFP (ab290), actin (ab6276), calnexin (ab22595) were from Abcam. Other antibodies were, phosphotyrosine (P5872, Merck), Rac1 (610650, BD Biosciences), TfR (13-6800, Thermo Fisher Scientific), Na/K-ATPase (NB300-146, Novus Biologicals) and Lamp1 (clone H4A3, Developmental Studies Hybridoma Bank). Secondary antibodies: anti-human unconjugated (A18819) anti-human Alexa488 (A11013), anti-human Alexa568 (A21090), anti-human Alexa647 (A21445), anti-rabbit Alexa488 (A11008) and anti-mouse Alexa488 (A11001) were from Thermo Fisher Scientific. HRP-conjugated secondary antibodies: anti-mouse (172–1011) anti-human (172–1033) anti-rabbit (172–1019) and anti-goat (172–1034) were from BioRad.
Soluble TfR ectodomain (aa 89–760) was purified from baculovirus-infected Sf9 cell culture supernatants. After equilibration to 20 mM Tris pH 8 and 500 mM NaCl and filtration through a 0.22-µm membrane, the protein was purified by Ni-affinity purification using a HisTrapExcel column (17-3712-06, GE Healthcare), followed by size exclusion chromatography using a Superdex200 16/60 column running in 20 mM HEPES pH 7.5, 150 mM NaCl.
Paul D., Stern O., Vallis Y., Dhillon J., Buchanan A, & McMahon H. (2023). Cell surface protein aggregation triggers endocytosis to maintain plasma membrane proteostasis. Nature Communications, 14, 947.
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Publication 2023
Actins Alexa Fluor 647 Antibodies Baculoviridae Biological Factors Biotin Calnexin Cell Culture Techniques CHO Cells Chromatography, Affinity Clone Cells Cloning Vectors Cytoplasm Dextran EGFR protein, human erbB-3, Proto-Oncogene Proteins ERBB2 protein, human Esters Filtration Fluorescein Gel Chromatography Goat HEPES High-Performance Liquid Chromatographies Homo sapiens Hybridomas Immunoglobulins Ligands lysosomal-associated membrane protein 1, human Mus Na(+)-K(+)-Exchanging ATPase Novus Phosphotyrosine Polyethyleneimine Proteins Rabbits SDS-PAGE Sodium Chloride Staphylococcal Protein A tetramethylrhodamine Tissue, Membrane TNFSF14 protein, human Transfection Transferrin Trastuzumab Tromethamine VAV1 protein, human VAV3 protein, human Wheat Germ Agglutinins
4
Inhibition of EGFR Phosphorylation in A431 Cells
The inhibition of EGFR phosphorylation was studied as described in the literature [13 (link)]. A431 cells (5 × 105 cells/well) were seeded in 6-well plates and incubated for 24 h. Then, the medium was replaced with fresh medium without fetal bovine serum. The next day, cells were incubated in the presence of the compound at the following concentrations: 0.1 nM, 1 nM, 10 nM, 50 nM, 100 nM, 1 μΜ, and 10 μΜ for 2 h, and then EGF was added (20 ng/mL) and left for 5 min. Two control samples were included in the experiment containing cell cultures without the compound and with or without EGF as a positive and negative control, respectively. Cells were lysed with cell lysis buffer (50 mM Tris-HCl pH 7.4, 150 mM, NaCl, 1 % Triton X-100, 0.25 % w/v sodium deoxycholate, 0.1 % w/v, sodium dodecyl sulfate, protease, and phosphatase inhibitors with EDTA). Ten micrograms of total protein for each sample were loaded onto polyacrylamide gel (8%) for electrophoresis. Subsequently, the proteins were separated by electrophoresis and transferred to a nitrocellulose membrane. Next, the membrane was washed three times in TBS-T buffer (tris-buffered saline and Tween 20), blocked for one hour in TBS-T with 5% milk (1% fat), and incubated for two hours minimum in a phosphotyrosine antibody (p-Tyr 100, mouse mAb) diluted 1/1000. Afterward, the membrane was washed three times in TBS-T and incubated in an anti-mouse IgG (HRP Linked) antibody diluted 1/1000 for 1 h. Then, the membrane was rewashed three times in TBS-T. The detection was performed using a chemiluminescent detection system (ECL kit, SignalFire ECL Reagent, Cell Signaling Technology, Auburn, MA, USA) according to the manufacturer’s instructions. Protein band quantification was performed using Image-J software, and the percentage of EGFR phosphorylation inhibition was calculated using GraphPad Prism 9.0 software.
Makrypidi K., Kiritsis C., Roupa I., Triantopoulou S., Shegani A., Paravatou-Petsotas M., Chiotellis A., Pelecanou M., Papadopoulos M, & Pirmettis I. (2023). Evaluation of Rhenium and Technetium-99m Complexes Bearing Quinazoline Derivatives as Potential EGFR Agents. Molecules, 28(4), 1786.
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Publication 2023
anti-IgG Buffers Cell Culture Techniques Cells Deoxycholic Acid, Monosodium Salt Edetic Acid EGFR protein, human Electrophoresis Fetal Bovine Serum Immunoglobulins inhibitors Milk, Cow's Mus Nitrocellulose Peptide Hydrolases Phosphoric Monoester Hydrolases Phosphorylation Phosphotyrosine polyacrylamide gels prisma Proteins Psychological Inhibition Saline Solution Sodium Chloride Sulfate, Sodium Dodecyl Tissue, Membrane Triton X-100 Tween 20
5
Benchmarking Signaling Network Interactions
stDPI, DPI and noDPI signaling networks were generated from U54-NET as described. Further, ground truth interactions were selected if they were identified as ST-K→S pairs by PDZ, SH3, WH1, and WW domain HSM analysis, since these represent the primary determinants of specific serine and threonine phosphopeptide interactions with ST-Ks. As a negative gold standard, we used candidate TK→S interactions with an inferred PTB, PTP and SH2 domain interaction with phosphotyrosine-containing peptide, since the used dataset for this benchmark (U54-NET), which is not enriched for phosphotyrosine peptides, should not be able to identify these interactions. This resulted in a context-specific reference dataset that separates between very likely direct and indirect interactions suitable for methodological comparisons. Receiver-Operating-Characteristics (ROC) curves were generated using the pROC R-package (version 1.17.0.1) and default parameters for each signaling network separately. P-values for ROC curve comparisons were also computed using pROC by DeLong’s test and using default parameters.
Rosenberger G., Li W., Turunen M., He J., Subramaniam P.S., Pampou S., Griffin A.T., Karan C., Kerwin P., Murray D., Honig B., Liu Y, & Califano A. (2023). Network-based elucidation of colon cancer drug resistance by phosphoproteomic time-series analysis. bioRxiv.
Publication Preprint 2023
Gold Peptides Phosphopeptides Phosphotyrosine Protein Domain Serine Threonine WW Domains

Top products related to «Phosphotyrosine»

1
Anti phosphotyrosine 4g10 by Merck Group
Sourced in United States, Germany
Anti-phosphotyrosine 4G10 is a laboratory reagent used for the detection and analysis of phosphorylated tyrosine residues in proteins. It is a monoclonal antibody that specifically binds to phosphorylated tyrosine residues, allowing for the identification and quantification of tyrosine phosphorylation events in protein samples.
2
Anti phosphotyrosine antibody 4g10 by Merck Group
Sourced in United States
The Anti-phosphotyrosine antibody (4G10) is a laboratory reagent used to detect and study tyrosine phosphorylation in proteins. It is a monoclonal antibody that specifically binds to phosphorylated tyrosine residues, a common post-translational modification in cellular signaling pathways. The 4G10 antibody can be used in various techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry, to identify and quantify phosphotyrosine-containing proteins.
3
Anti phosphotyrosine by Merck Group
Sourced in France, United States
Anti-phosphotyrosine is a lab equipment product used to detect and quantify the presence of phosphorylated tyrosine residues in proteins. It functions by binding to and identifying phosphorylated tyrosine groups within a sample.
4
Pvdf membrane by Merck Group
Sourced in United States, Germany, China, United Kingdom, Morocco, Ireland, France, Italy, Japan, Canada, Spain, Switzerland, New Zealand, India, Hong Kong, Sao Tome and Principe, Sweden, Netherlands, Australia, Belgium, Austria
PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
5
Imagequant software by GE Healthcare
Sourced in United States, United Kingdom, Sweden, Germany, Austria, Australia
ImageQuant software is a data analysis tool designed for quantitative analysis of digital images. It enables users to capture, process, and analyze images from various laboratory equipment, such as gel imagers and blot imaging systems. The software provides tools for image acquisition, image processing, and data analysis, allowing researchers to quantify and compare signals within their samples.
6
Bovine serum albumin (bsa) by Merck Group
Sourced in United States, Germany, United Kingdom, China, Italy, Japan, France, Sao Tome and Principe, Canada, Macao, Spain, Switzerland, Australia, India, Israel, Belgium, Poland, Sweden, Denmark, Ireland, Hungary, Netherlands, Czechia, Brazil, Austria, Singapore, Portugal, Panama, Chile, Senegal, Morocco, Slovenia, New Zealand, Finland, Thailand, Uruguay, Argentina, Saudi Arabia, Romania, Greece, Mexico
Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.
7
Phosphorimager screen by GE Healthcare
Sourced in United States, United Kingdom
The PhosphorImager screen is a laboratory equipment used for the detection and quantification of radioactively labeled samples. It functions by capturing the luminescence emitted from phosphor-coated screens that have been exposed to radioactive samples, allowing for the visualization and analysis of these samples.
8
Lipofectamine 2000 by Thermo Fisher Scientific
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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
9
Phosphotyrosine 4g10 by Merck Group
Sourced in United States
Phosphotyrosine (4G10) is a monoclonal antibody that specifically recognizes phosphorylated tyrosine residues. It is a widely used tool in molecular biology and biochemistry research for the detection and analysis of tyrosine phosphorylation events.
10
Typhoon 8600 by GE Healthcare
Sourced in United States, United Kingdom
The Typhoon 8600 is a high-performance imager designed for a variety of life science applications. It can capture images of gels, blots, and other samples using multiple detection modes, including fluorescence, chemiluminescence, and phosphor imaging.

More about "Phosphotyrosine"

Phosphotyrosine, also known as pTyr or p-Tyr, is a key post-translational modification (PTM) where a phosphate group is covalently attached to the hydroxyl group of the amino acid tyrosine.
This modification plays a crucial role in cellular signaling pathways, regulating protein-protein interactions, enzyme activity, and subcellular localization of proteins.
Phosphotyrosine-containing proteins are involved in a wide range of biological processes, such as cell growth, differentiation, metabolism, and immune response.
Researchers studying phosphotyrosine-mediated signaling can leverage powerful tools like Anti-phosphotyrosine 4G10 antibody, which specifically recognizes and binds to phosphorylated tyrosine residues.
The 4G10 antibody can be used in techniques like Western blotting, immunoprecipitation, and immunofluorescence to detect and analyze phosphotyrosine-containing proteins.
PVDF membranes are commonly used in Western blotting to immobilize and detect phosphotyrosine proteins.
The ImageQuant software can be utilized to quantify and analyze the signal intensity of phosphotyrosine bands on the blots.
Additionally, the PhosphorImager screen can be used to visualize and quantify radioactively labeled phosphotyrosine-containing proteins.
To study phosphotyrosine signaling in cells, researchers may employ transfection reagents like Lipofectamine 2000 to introduce plasmids or siRNAs targeting phosphotyrosine-related genes.
Bovine serum albumin (BSA) is often used as a blocking agent to reduce non-specific binding in immunoassays.
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