Phospho met tyr1234 1235

Manufactured by Cell Signaling Technology

Sourced in United States

Phospho-Met (Tyr1234/1235) is a lab equipment product that detects phosphorylation of the Met (receptor tyrosine kinase) protein at tyrosine residues 1234 and 1235. This product is used to study cell signaling pathways involving the Met protein.

Automatically generated - may contain errors

Lab products found in correlation

9 protocols using phospho met tyr1234 1235

Targeted Protein Pathway Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Erlotinib and osimertinib were purchased from Selleck Chemicals (Houston, TX, USA). Crizotinib was from Sigma-Aldrich (St. Louis, MO, USA). Chloroquine was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The primary antibodies used for immunoblot analyses were the products of Cell Signaling Technology (Danvers, MA, USA); anti-eIF3c (#2068), EGFR (#4267), phospho-EGFR (Tyr1068) (#3777), Akt (#9272), phospho-Akt (Ser473) (#9271), ERK1/2 (#9102), phospho-ERK1/2 (Thr202/Tyr204) (#9101), Met (#8198), phospho-Met (Tyr1234/1235) (#3077), LC3B (#3868), and β-actin (#4970).

Shintani T., Higashisaka K., Maeda M., Hamada M., Tsuji R., Kurihara K., Kashiwagi Y., Sato A., Obana M., Yamamoto A., Kawasaki K., Lin Y., Kijima T., Kinehara Y., Miwa Y., Maeda S., Morii E., Kumanogoh A., Tsutsumi Y., Nagatomo I, & Fujio Y. (2018). Eukaryotic translation initiation factor 3 subunit C is associated with acquired resistance to erlotinib in non-small cell lung cancer. Oncotarget, 9(101), 37520-37533.

+ Open protocol

+ Expand

Immunoblotting Analysis of Signaling Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ascites cells were lysed in buffer containing 20 mM Tris (pH 7.5), 150 mM NaCl, 1% NP40, 1 mM EDTA, 5 mM NaF and 5 mM Na₃VO₄. Solid tumor tissues were homogenized using the microtube pestles (Thomas Scientific) prior to add the lysis buffer above. Total cell lysates (30 µg) were resolved by 10% SDS–PAGE, transferred to nitrocellulose membranes, and incubated with 100–200 µg/mL of the following antibodies: phospho-Akt (Ser473), phospho-p44/42 ERK (Thr202/Tyr204), phospho-NF-κB p65 (Ser536), phospho-mTOR (Ser2448), phospho-B-Raf (Ser445), phospho-p38 MAPK (Thr180/Tyr182), phospho-Met (Tyr1234/1235) (Cell Signaling Technologies), KSHV-LANA (ABI) and K8.1A (Thermo Scientific-Pierce, Clone: 8C12G10G1). For loading controls, lysates were also incubated with antibodies detecting β-Actin (Sigma). Immunoreactive bands were developed using an enhanced chemiluminescence reaction (Perkin-Elmer).

Dai L., Trillo-Tinoco J., Bai L., Kang B., Xu Z., Wen X., Valle L.D, & Qin Z. (2014). Systematic Analysis of a Xenograft Mice Model for KSHV+ Primary Effusion Lymphoma (PEL). PLoS ONE, 9(2), e90349.

+ Open protocol

+ Expand

Immunohistochemical Evaluation of Tumor Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemistry was performed using formalin‐fixed paraffin‐embedded sections. Following deparaffinization with xylene and rehydration with graded ethanol, antigen retrieval was performed using 0.09% (v/v) unmasking solution (Vector Labs) for 30 min in a steamer. Inactivation of endogenous peroxidases was carried out using 3% hydrogen peroxide (Sigma) for 10 min. Secondary antibody staining and biotin–streptavidin incubation were performed using species‐specific VECTASTAIN Elite ABC Kits (Vector Labs). DAB Peroxidase Substrate Kit (Vector Labs) was utilized for antibody detection. Primary antibodies used were anti‐pH3 Ser10 (1:200, Cell Signalling, 9701), cleaved caspase 3 (1:200, Cell Signalling, 9661) and Phospho‐Met (Tyr1234/1235) (1:300, Cell Signaling, 3077). Tumour sections were visualized under a TissueFAXS slide scanning platform (TissueGnostics, Vienna, Austria). For the pH3 and Casp3, the quantitation was performed using StrataQuest software (TissueGnostics) to determine the percentage of pH3⁺ or Casp3⁺ cells.

Salgueiro L., Buccitelli C., Rowald K., Somogyi K., Kandala S., Korbel J.O, & Sotillo R. (2020). Acquisition of chromosome instability is a mechanism to evade oncogene addiction. EMBO Molecular Medicine, 12(3), e10941.

+ Open protocol

+ Expand

Analysis of AXL and Met Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies against AXL (#8661), phospho-AXL (Tyr702) (#5724), phospho-MET (Tyr1234/1235) (#3077), AKT (#4685), phospho-AKT (ser473) (#4058), ERK (#4696), phospho-ERK (Thr202/Tyr204) (4376), GSK3β (9832), phospho-GSK3β (Ser9) (#5558), β-catenin (#8480), Snail (#3879), phospho-Src-Y416 (#6943) were purchased from Cell Signaling Technology. Antibody against Met (sc-161-R) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Antibody against AXL (#AF154) purchased from R&D systems (Minneapolis, MN) was used for IHC. Antibodies against VEGF (ab150766) and CD31 (ab28364) were purchased from Abcam (Cambridge, MA). Recombinant GAS6 (#885-GS-050) was purchased from R&D systems (Minneapolis, MN). Recombinant HGF (#GF116) was purchased from Millipore. Mitomycin C (#475820) was purchased from Calbiochem (Billerica, MA). Sunitinib (#s1042) was purchased from Selleckchem (Houston, TX). Cabozantinib was provided by Exelixis (South San Francisco, CA).

Zhou L., Liu X.D., Sun M., Zhang X., German P., Bai S., Ding Z., Tannir N., Wood C.G., Matin S.F., Karam J.A., Tamboli P., Sircar K., Rao P., Rankin E.B., Laird D.A., Hoang A.G., Walker C.L., Giaccia A.J, & Jonasch E. (2015). Targeting MET and AXL overcomes resistance to sunitinib therapy in renal cell carcinoma. Oncogene, 35(21), 2687-2697.

+ Open protocol

+ Expand

Comprehensive Antibody Profiling Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Monoclonal antibodies used in this study were directed against phospho-MET (Tyr1234/1235) (Cell Signaling Technology), MET (clone D1C2, Cell Signaling Technology), phospho-EGFR (Tyr845) (Cell Signaling Technology), phospho-ALK (Tyr1064) (Cell Signaling Technology), phospho-ATM (Ser1981) (Cell Signaling Technology), phospho-c-Raf (Ser259) (Cell Signaling Technology), phospho-AKT (Ser473) (Cell Signaling Technology), phospho-MAPK (Thr202/Tyr204) (Cell Signaling Technology), phospho-H2AX (Ser139) (Upstate), Ki67 (Cell Signaling Technology) and ß-actin (Millipore Chemicon).

Orlando E., Medo M., Bensimon A., Quintin A., Riedo R., Roth S.M., Riether C., Marti T.M., Aebersold D.M., Medová M., Aebersold R, & Zimmer Y. (2022). An oncogene addiction phosphorylation signature and its derived scores inform tumor responsiveness to targeted therapies. Cellular and Molecular Life Sciences, 80(1), 6.

+ Open protocol

+ Expand

Western Blot Analysis of Hepatic Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hepatic tissues from individual mice were lysed in radio immune precipitation buffer (50 mmol/L Tris‐HCl, 150 mmol/L NaCl, 5 mmol/L MgCl2, 2 mmol/L ethylenediaminetetraacetic acid, 1 mmol/L NaF, 1% NP40 and 0.1% sodium dodecyl sulfate). Western blotting was measured as previously described13 and was evaluated with antibodies against farnesoid X receptor (FXR; 1:3000, ab126602; Abcam Inc., Cambridge, MA, USA), short heterodimer partner (SHP; 1:1000, sc‐30169; Santa Cruz Biotech, Santa Cruz, CA, USA), fatty acid synthase (FAS; 1:1000, 610962; BD Inc., San Jose, CA, USA), sterol regulatory element‐binding factor‐1c (SREBP‐1c; 1:1000, sc‐8984; Santa Cruz Biotech), peroxidase proliferator‐activated receptor‐α (PPARα; 1:1000, ab8934; Abcam Inc.), Phospho‐Met (Tyr1234/1235; #3126, Cell Signaling Technology Inc.), Met (C‐28; 1:1000, sc‐161; Santa Cruz Biotech) and GAPDH (Cell Signaling Technology Inc.). Protein expression levels were quantitated using the Image J software (https://imagej.nih.gov/ij/).

Jing Y., Sun Q., Xiong X., Meng R., Tang S., Cao S., Bi Y, & Zhu D. (2018). Hepatocyte growth factor alleviates hepatic insulin resistance and lipid accumulation in high‐fat diet‐fed mice. Journal of Diabetes Investigation, 10(2), 251-260.

+ Open protocol

+ Expand

Protein Extraction from Mouse Brain and Cell Cultures

Check if the same lab product or an alternative is used in the 5 most similar protocols

Brains isolated from mock-irradiated or irradiated C57BL/6J mice were homogenized in NP-40 lysis buffer (10 mM Tris–HCl, pH 8.0, 150 mM NaCl, 1% NP-40, and protease and phosphatase inhibitor cocktails). Extracts were clarified by centrifugation at 16,000 g at 4 °C and the supernatant was collected and processed for western blotting. Cells in culture were washed with cold PBS, collected using a cell scraper, and centrifuged at 2000 g for 5 min. The pellets were lysed with NP-40 lysis buffer, followed by sonication and centrifugation at 16,000 g at 4°C. The supernatant was collected, and protein concentration was measured by the Bradford assay (Bio-Rad) prior to preparation of samples for SDS-PAGE and western blotting. Antibodies used for western blotting are as follows: HGF (Abcam, Cat. No. 83760), p21 (Abcam, Cat. No. 1888224), β-Actin (Santa Cruz, Cat. No. 47778), Lamin B1 (Abcam, Cat. No. 16048), phospho-Met Tyr1234/1235 (Cell Signaling, Cat. No. 3077S), Met (Cell Signaling, Cat. No. 8198), GAPDH (Cell Signaling, Cat. No. 5174), and HRP-conjugated secondary antibodies (Bio-Rad).

Fletcher-Sananikone E., Kanji S., Tomimatsu N., Cristofaro L.F., Kollipara R.K., Saha D., Floyd J.R., Sung P., Hromas R., Burns T.C., Kittler R., Habib A.A., Mukherjee B, & Burma S. (2021). Elimination of radiation-induced senescence in the brain tumor microenvironment attenuates glioblastoma recurrence. Cancer research, 81(23), 5935-5947.

+ Open protocol

+ Expand

Comprehensive Signaling Pathway Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Phospho-Met (Tyr1349), Phospho-Met (Tyr1003), phospho-Rac1/cdc42(Ser71), Rac1/2/3, phospho-p44/42 MAPK (Erk1/2), phospho-STAT3 (Tyr705), STAT3, Ki-67, p44/42 MAPK (Erk1/2), phospho-PDK1 (Ser241), PDK1, PTEN, PI3 kinase p110α antibody, phospho-PI3 kinase p85 (Tyr458)/p55 (Tyr199), phospho-Akt (Ser473), phospho-Akt (Thr308), phospho-mTOR (Ser2448), mTOR, phospho-GSK-3α/β (Ser21/9), GSK-3β (3D10), phospho-p70S6 kinase (T389), p70S6 kinase, phospho-4E-BP1 (Thr37/46) (236B4), 4E-BP1 (53H11), phospho-eI4FE (Ser209), eI4FE, phospho-Cyclin D1, Cyclin D1, Cyclin E1 (HE12), Cyclin A, PCNA, phospho-Rb (S807/811), Rb, Caspase-3 (8G10), Caspase-9, PARP, Bad, Cleaved caspase-3 (Asp175) (5A1E) and Phospho-Met (Tyr1234/1235) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). The Met kinase assay/inhibitor screening kit (# KA0055) was purchased from Abnova (Walnut, CA, USA).

Fan Y., Ding H., Kim D., Bach D.H., Hong J.Y., Xu Y, & Lee S.K. (2019). Antitumor Activity of DFX117 by Dual Inhibition of c-Met and PI3Kα in Non-Small Cell Lung Cancer. Cancers, 11(5), 627.

+ Open protocol

+ Expand

Antibody Characterization for CD44v6 Targeting

Check if the same lab product or an alternative is used in the 5 most similar protocols

The antibodies used in the present study were directed against rat CD44v6 (1.1ASML, described in [20 (link)]) and human CD44v6 (VFF18; Boehringer Ingelheim), Erk-1 (K-23; Santa Cruz), phospho-p44/42 MAPK (Thr202/Tyr204; Cell Signaling Technology), penta-His (Qiagen), human HGF (AF-294-NA; R&D Systems), rat Met (B2; Santa Cruz), human Met (25H2; Cell Signaling Technology) phospho-Met (Tyr1234/1235; Cell Signaling Technology), human TGF-α (transforming growth factor alpha; Peprotech), VEGFR-2 (A3; Santa Cruz) and human VEGF-A165 (VEGF₁₆₅) (AF-293-NA; R&D Systems). The secondary antibodies labelled with horseradish peroxidase (HRP) were from Dako, the phycoerythrin (PE)-labelled antibody from Thermo Scientific and the FITC–streptavidin from Invitrogen. The human HGF was a generous gift from Ermanno Gherardi (University of Pavia, Pavia, Italy). Human VEGF₁₆₅ and human TGF-α were from Peprotech. The peptides have already been described in [14 (link)] and their sequences are: rat CD44v6 5mer–NEWQG, rat CD44v6 14mer–KEKWFENEWQGKNP, human CD44v6 5mer–NRWHE, human CD44v6 14mer–KEQWFGNRWHEGYR, mouse CD44v6 5mer–NGWQG and mouse CD44v6 14mer–QETWFQNGWQGKNP.

Volz Y., Koschut D., Matzke-Ogi A., Dietz M.S., Karathanasis C., Richert L., Wagner M.G., Mély Y., Heilemann M., Niemann H.H, & Orian-Rousseau V. (2015). Direct binding of hepatocyte growth factor and vascular endothelial growth factor to CD44v6. Bioscience Reports, 35(4), e00236.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!