py527 cell signaling  (Cell Signaling Technology Inc)

Journal: The American Journal of Pathology

Article Title: Src Kinase Is Biphosphorylated at Y416/Y527 and Activates the CUB-Domain Containing Protein 1/Protein Kinase C δ Pathway in a Subset of Triple-Negative Breast Cancers

doi: 10.1016/j.ajpath.2019.10.017

Figure Lengend Snippet: Antibodies and Dilutions Used for IHC and Western Blot Analysis

Article Snippet: Antibody dilutions for each method are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antigen Antibody information, manufacturer/catalog no. Western blot dilution IHC dilution Antigen retrieval ∗ FOXA1 Abcam/ab173287 NA 1:4000 1 Vimentin Agilent/M0725 NA 1:2000 2 Sox10 Bio SB/BSB2583 NA 1:100 1 CK5/6 Thermo Fisher Scientific/180267 NA 1:50 2 CDCP1 Cell Signaling Technology/4115 1:1000 1:100 1 CDCP1_pY743 Cell Signaling Technology/14965 1:1000 1:800 1 CDCP1_pY734 Cell Signaling Technology/9050 1:1000 NA NA Src Cell Signaling Technology/2108 1:1000 NA NA Lck Cell Signaling Technology/2752 1:1000 NA NA Lyn Cell Signaling Technology/2732 1:1000 NA NA SFK_pY416 Cell Signaling Technology/2101 1:1000 1:200 1 SFK_pY527 Cell Signaling Technology/2105 1:1000 NA NA PKCδ Cell Signaling Technology/2058 1:1000 NA NA PKCδ_pY311 Abcam/ab76181 1:2500 1:3000 1 N-cadherin Cell Signaling Technology/13116 NA 1:200 1 Slug Cell Signaling Technology/9585 NA 1:100 1 AMPKα Cell Signaling Technology/2532 1:1000 NA NA AMPKα_pT172 Cell Signaling Technology/2535 1:1000 NA NA β-Actin Leica Biosystems/A5441 1:5000 NA NA CD3 Novocastra/NCL-L-CD3-565 NA 1:50 2 PD1 Cell Signaling Technology/86163 NA 1:200 2 PD-L1 Cell Signaling Technology/13684 NA 1:400 2 CTLA4 Santa Cruz Biotechnology/sc-376016 NA 1:400 2 Open in a separate window AMPK, AMP-activated protein kinase; CDCP, CUB-domain containing protein; CK5/6, cytokeratin 5/6; CTLA4, cytotoxic T-lymphocyte protein 4; FOXA1, forkhead box protein A1; IHC, immunohistochemistry; Lck, tyrosine-protein kinase Lck; Lyn, tyrosine-protein kinase Lyn; NA, not applicable; PD1, programmed cell death protein 1; PD-L1, programmed cell death 1 ligand 1; PKC, protein kinase C; Sox10, Sry-box transcription factor 10.

Techniques: Western Blot

Journal:

Article Title: CHARACTERIZATION OF NEURONAL SRC KINASE PURIFIED FROM A BACTERIAL EXPRESSION SYSTEM

doi: 10.1016/j.pep.2010.06.004

Figure Lengend Snippet: A: purified Src proteins expressed in E. coli BL21(DE3) Cells. Cms: visualized with Coomassie G250. The numbers on the left of the gel indicate the molecular mass (kDa). WB: Western blotting with antibodies as indicated. SrcpY535 (corresponding to c-SrcpY527): probed with anti-pY527 antibody (Cell Signaling); SrcpY424 (corresponding to c-SrcpY416): probed with anti-pY416 antibody (Cell signaling). Src: an antibody recognizing the C-tail of Src (Upstate Biotechnology). B: the kinetics of kinase activity of purified n-Src proteins. Values in brackets indicate number of experimental repeats.

Article Snippet: Antibodies including anti-Src antibody (Millipore), anti-pY527 antibody (Cell Signaling), anti-pY416 antibody (Cell Signaling), and anti-phosphotyrosine antibody (4G10, Upstate) were used.

Techniques: Purification, Western Blot, Activity Assay