Pshp2 y542

Manufactured by Abcam

PShp2 Y542 is a recombinant protein that represents a specific phosphorylation site of the SHP2 protein. It can be used as a research tool to study the role and regulation of SHP2 phosphorylation.

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Lab products found in correlation

α sma, by Cell Signaling Technology (2 mentions) Monoclonal flag m2, by Merck Group (1 mentions) P mek1 2, by Santa Cruz Biotechnology (1 mentions) Vinculin v9264, by Merck Group (1 mentions) Caspase 9, by Cell Signaling Technology (1 mentions) Cleaved parp, by Cell Signaling Technology (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Cleaved caspase 9, by Cell Signaling Technology (1 mentions) Active β catenin, by Cell Signaling Technology (1 mentions) Ve cadherin, by Thermo Fisher Scientific (1 mentions) Bicinchoninic acid assay, by Beyotime (1 mentions) Lamin b1, by Proteintech (1 mentions) β tubulin, by Huabio (1 mentions) β actin, by Huabio (1 mentions) Nitrocellulose membrane, by Pall Corporation (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Ripa buffer, by Beyotime (1 mentions) Phosstop, by Roche (1 mentions) β catenin, by Abcam (1 mentions) Citrate antigen retrieval solution, by Solarbio (1 mentions)

3 protocols using pshp2 y542

Immunoblotting Antibody Validation Protocol

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Rabbit polyclonal antibodies against Src (#2109, 1:5000), phosphorylated (p)Src (#2101, 1:1000), pAKT (#9271, 1:1000), AKT (#9272, 1:1000), ERK (#9102, 1:1000), SYK (#2712, 1:1000), FAK (#3285, 1:1000), pTyr (P-Tyr-1000) (#8954, 1:2000), cleaved caspase-3 (#9664, 1:1000), cleaved PARP (#9541, 1:1000), PARP (#9542, 1:1000), cleaved caspase-9 (#9661, 1:1000), caspase-9 (#9508, 1:1000), and HA (#3724, 1:5000) were obtained from Cell Signaling Technologies. Polyclonal IgG (sc-2027), pERK (sc-7383, 1:500), CBL (sc-170, 1:500), SHP2 (sc-280, 1:1000), pMEK1/2 (sc-81503, 1:500), and MEK-1 (sc-6250, 1:500) were obtained from Santa Cruz Biotechnology. p-SHP2(Y542) (ab62322, 1:20,000) was obtained from Abcam. Monoclonal antibodies against Pan-Ras (OP40, 1:500), HA (12CA5, 1:500), and pTyr (4G10) (05–321, 1:1000) were obtained from Boehringer Ingelheim and Millipore, respectively. Monoclonal FLAG-M2 (F1804, 1:2000) and Vinculin (V9264, 1:2000) were obtained from Sigma.

Kano Y., Gebregiworgis T., Marshall C.B., Radulovich N., Poon B.P., St-Germain J., Cook J.D., Valencia-Sama I., Grant B.M., Herrera S.G., Miao J., Raught B., Irwin M.S., Lee J.E., Yeh J.J., Zhang Z.Y., Tsao M.S., Ikura M, & Ohh M. (2019). Tyrosyl phosphorylation of KRAS stalls GTPase cycle via alteration of switch I and II conformation. Nature Communications, 10, 224.

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Western Blot Analysis of Cell Signaling

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Cells or lung tissue were lysed in RIPA buffer (Beyotime) containing Na₃VO₄ (1 mM), protease inhibitor cocktail, and phosphatase inhibitor (phosSTOP, Roche), and the protein concentration was quantified by bicinchoninic acid assay (Beyotime). Equal amounts of protein were separated by SDS-PAGE, then transferred onto a nitrocellulose membrane (Pall Corp). The membranes were incubated at 4°C overnight with primary antibodies against Jag1 (Cell Signaling Technology), Shp2 (Cell Signaling Technology), pShp2 Y542 (Abcam), β-catenin (Abcam), active-β-catenin (Cell Signaling Technology), Col1α (Proteintech Group), α-SMA (Cell Signaling Technology), CD31 (Abcam), VE-cadherin (Ebioscience), Lamin-B1 (Proteintech Group), GAPDH (Daige), β-tubulin (Huabio), and β-actin (Huabio). The matched fluorescein-linked secondary antibodies (LI-COR Biosciences) were used to visualize proteins by incubation at room temperature for 1 h. The membranes were scanned with Odyssey (LI-COR Biosciences) and quantified by ImageJ.

Liu P., Li Y., Li M., Zhou H., Zhang H., Zhang Y., Xu J., Xu Y., Zhang J., Xia B., Cheng H., Ke Y, & Zhang X. (2022). Endothelial Shp2 deficiency controls alternative activation of macrophage preventing radiation-induced lung injury through notch signaling. iScience, 25(3), 103867.

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Immunohistochemical Analysis of Lung Tissue

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Lung sections were incubated at 65°C for 1 h, then heated in citrate antigen-retrieval solution for 30 min (Solarbio). 0.5% TritonX-100 was used for permeabilization and 5% FBS was used for blocking. Sections were incubated with primary antibodies against Jag1 (Cell Signaling Technology), Shp2 (Sigma-Aldrich), pShp2 Y542 (Abcam), IB4 (Invitrogen), α-SMA (Cell Signaling Technology), Arg1 (Proteintech Group), and CD68 (Ebioscience) overnight at 4°C. The matched fluorescein-linked secondary antibodies (Invitrogen) were used for visualization by incubation at room temperature for 1 h. Nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI, Beyotime). Images were acquired with an Olympus IX81-FV1000 microscope.
Cells on slides were fixed with 4% paraformaldehyde for 20 min, then permeabilized with 0.1% TritonX-100 for 20 min.

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