Marker:Article Title: Structural analysis of TRIM family PRYSPRY domains and its implications for E3-ligand design
Article Snippet: .. Marker proteins used: bovine serum albumin (MW = 66 kDa; Sigma-Aldrich), hen egg white ovalbumin (MW = 44 kDa, Sigma-Aldrich), bovine erythrocyte carbonic anhydrase (MW = 30 kDa, MP Biomedicals), bovine ribonuclease A (MW = 13.7 kDa, Sigma-Aldrich). ..
other:Article Title: Streamlining Peptide Mapping LC-MS Approach for Studying Fusion Peptide-Conjugated Vaccine Immunogens
Article Snippet: Carbonic anhydrase (CA) and CRM197 were obtained from MP Biomedical (Irvine, CA) and Pfenex Inc. (San Diego, CA), respectively.
Recombinant:Article Title: Multiprobe Photoproximity Labeling of the EGFR Interactome in Glioblastoma Using Red-Light
Article Snippet: Duolink® Proximity Ligation Assay kits (Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, DUO92002; Duolink® In Situ PLA® Probe Anti-Mouse MINUS, DUO92004; Duolink® In Situ Detection Reagents Orange, DUO92007) were purchased from EMD Millipore. .. Structure of probes used in this study HN NH S O N H O NH2 H H O N H HN NH S O OH H H O N H HN NH S O H NO O O O H H CF3 NN O N H HN NH S O N H O O O H H O N3 O H N NH2NH O HN NH S O H H Dz-Bt Az-Bt Hy-BtPhenol-Bt An-Bt Biotin-Phenol (Biotinyl tyramide) was purchased from SIGMA (SML2135); Biotin LC hydrazide was purchased from Santa Cruz Biotechnology (sc-257155); Biotin-aniline was purchased from SIGMA (SML2946); Biotin-PEG3-Ph(4-N3) was purchased from Iris Biotech GMBH (PEG6795); Biotin-PEG3Diazrine was prepared accoriding to the literature.1 Recombinant protein used in this study Carbonic anhydrase (CA) was purchased from Mp Biomedicals Inc (ICN15387925), Bovine serum albumin (BSA) was purchased from SIGMA (A9647); Recombinant Human Epidermal growth factor (EGF) was purchased from Thermo Fisher scientific (AF-100-15-1mg) Antibodies used in this study Streptavidin HRP-Conjugated Streptavidin , Thermo Fisher scientific, N100 1:1000 in TBST Western Blot Streptavidin IRDye® 800CW Streptavidin Li Cor Inc, 926-32230 1:1000 in TBST Western Blot Streptavidin IRDye® 680LT Streptavidin 1:1000 in TBST 34 Li Cor Inc, 926-68031 Western Blot Streptavidin DyLightTM 550, Thermo Fisher scientific, 84542 1:1000 in DPBS Flow Cyctometry Goat anti-Rabbit IRDye® 800CW, 926-32211 1:10000 in TBSTWestern Blot Cetuximab (anti-EGFR) Selleck Chemicals LLC, A2000 Anti-EGFR Cell Signaling Technology, 4267S 1:500 in TBSTWestern Blot Human IgG Thermo Fisher scientific, 0931431 Anti-EGFR BD PharmingenTM, 555330 1:200 in Proximity Ligation Assay Anti-NEO1 Thermo Fisher scientific, 20246-1-AP 1:200 in Proximity Ligation Assay Goat anti-Human-IgG 647 Thermo Fisher scientific, A21445 1:1000 in DPBSFlow Cyctometry Anti-β-actin Cell Signaling Technology, 3700 1:500 in TBSTWestern Blot Goat anti-Mice IRDye® 680CW, 926-68070 1:10000 in TBSTWestern Blot Cell Culture HEK 293T cells (ATCC CRL-3216) and A549 cells (ATCC CCL-185) were cultured as a monolayer in DMEM (Gibco, 10566016), supplemented with 10% v/v FBS (Thermo Fisher), 100 U ml-1 penicillin (Thermo Fisher), and 100 μg ml-1 streptomycin (Thermo Fisher) on 15 cm Nunclon Delta dishes (Thermo Fisher). ..
Western Blot:Article Title: Multiprobe Photoproximity Labeling of the EGFR Interactome in Glioblastoma Using Red-Light
Article Snippet: Duolink® Proximity Ligation Assay kits (Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, DUO92002; Duolink® In Situ PLA® Probe Anti-Mouse MINUS, DUO92004; Duolink® In Situ Detection Reagents Orange, DUO92007) were purchased from EMD Millipore. .. Structure of probes used in this study HN NH S O N H O NH2 H H O N H HN NH S O OH H H O N H HN NH S O H NO O O O H H CF3 NN O N H HN NH S O N H O O O H H O N3 O H N NH2NH O HN NH S O H H Dz-Bt Az-Bt Hy-BtPhenol-Bt An-Bt Biotin-Phenol (Biotinyl tyramide) was purchased from SIGMA (SML2135); Biotin LC hydrazide was purchased from Santa Cruz Biotechnology (sc-257155); Biotin-aniline was purchased from SIGMA (SML2946); Biotin-PEG3-Ph(4-N3) was purchased from Iris Biotech GMBH (PEG6795); Biotin-PEG3Diazrine was prepared accoriding to the literature.1 Recombinant protein used in this study Carbonic anhydrase (CA) was purchased from Mp Biomedicals Inc (ICN15387925), Bovine serum albumin (BSA) was purchased from SIGMA (A9647); Recombinant Human Epidermal growth factor (EGF) was purchased from Thermo Fisher scientific (AF-100-15-1mg) Antibodies used in this study Streptavidin HRP-Conjugated Streptavidin , Thermo Fisher scientific, N100 1:1000 in TBST Western Blot Streptavidin IRDye® 800CW Streptavidin Li Cor Inc, 926-32230 1:1000 in TBST Western Blot Streptavidin IRDye® 680LT Streptavidin 1:1000 in TBST 34 Li Cor Inc, 926-68031 Western Blot Streptavidin DyLightTM 550, Thermo Fisher scientific, 84542 1:1000 in DPBS Flow Cyctometry Goat anti-Rabbit IRDye® 800CW, 926-32211 1:10000 in TBSTWestern Blot Cetuximab (anti-EGFR) Selleck Chemicals LLC, A2000 Anti-EGFR Cell Signaling Technology, 4267S 1:500 in TBSTWestern Blot Human IgG Thermo Fisher scientific, 0931431 Anti-EGFR BD PharmingenTM, 555330 1:200 in Proximity Ligation Assay Anti-NEO1 Thermo Fisher scientific, 20246-1-AP 1:200 in Proximity Ligation Assay Goat anti-Human-IgG 647 Thermo Fisher scientific, A21445 1:1000 in DPBSFlow Cyctometry Anti-β-actin Cell Signaling Technology, 3700 1:500 in TBSTWestern Blot Goat anti-Mice IRDye® 680CW, 926-68070 1:10000 in TBSTWestern Blot Cell Culture HEK 293T cells (ATCC CRL-3216) and A549 cells (ATCC CCL-185) were cultured as a monolayer in DMEM (Gibco, 10566016), supplemented with 10% v/v FBS (Thermo Fisher), 100 U ml-1 penicillin (Thermo Fisher), and 100 μg ml-1 streptomycin (Thermo Fisher) on 15 cm Nunclon Delta dishes (Thermo Fisher). ..
Proximity Ligation Assay:Article Title: Multiprobe Photoproximity Labeling of the EGFR Interactome in Glioblastoma Using Red-Light
Article Snippet: Duolink® Proximity Ligation Assay kits (Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, DUO92002; Duolink® In Situ PLA® Probe Anti-Mouse MINUS, DUO92004; Duolink® In Situ Detection Reagents Orange, DUO92007) were purchased from EMD Millipore. .. Structure of probes used in this study HN NH S O N H O NH2 H H O N H HN NH S O OH H H O N H HN NH S O H NO O O O H H CF3 NN O N H HN NH S O N H O O O H H O N3 O H N NH2NH O HN NH S O H H Dz-Bt Az-Bt Hy-BtPhenol-Bt An-Bt Biotin-Phenol (Biotinyl tyramide) was purchased from SIGMA (SML2135); Biotin LC hydrazide was purchased from Santa Cruz Biotechnology (sc-257155); Biotin-aniline was purchased from SIGMA (SML2946); Biotin-PEG3-Ph(4-N3) was purchased from Iris Biotech GMBH (PEG6795); Biotin-PEG3Diazrine was prepared accoriding to the literature.1 Recombinant protein used in this study Carbonic anhydrase (CA) was purchased from Mp Biomedicals Inc (ICN15387925), Bovine serum albumin (BSA) was purchased from SIGMA (A9647); Recombinant Human Epidermal growth factor (EGF) was purchased from Thermo Fisher scientific (AF-100-15-1mg) Antibodies used in this study Streptavidin HRP-Conjugated Streptavidin , Thermo Fisher scientific, N100 1:1000 in TBST Western Blot Streptavidin IRDye® 800CW Streptavidin Li Cor Inc, 926-32230 1:1000 in TBST Western Blot Streptavidin IRDye® 680LT Streptavidin 1:1000 in TBST 34 Li Cor Inc, 926-68031 Western Blot Streptavidin DyLightTM 550, Thermo Fisher scientific, 84542 1:1000 in DPBS Flow Cyctometry Goat anti-Rabbit IRDye® 800CW, 926-32211 1:10000 in TBSTWestern Blot Cetuximab (anti-EGFR) Selleck Chemicals LLC, A2000 Anti-EGFR Cell Signaling Technology, 4267S 1:500 in TBSTWestern Blot Human IgG Thermo Fisher scientific, 0931431 Anti-EGFR BD PharmingenTM, 555330 1:200 in Proximity Ligation Assay Anti-NEO1 Thermo Fisher scientific, 20246-1-AP 1:200 in Proximity Ligation Assay Goat anti-Human-IgG 647 Thermo Fisher scientific, A21445 1:1000 in DPBSFlow Cyctometry Anti-β-actin Cell Signaling Technology, 3700 1:500 in TBSTWestern Blot Goat anti-Mice IRDye® 680CW, 926-68070 1:10000 in TBSTWestern Blot Cell Culture HEK 293T cells (ATCC CRL-3216) and A549 cells (ATCC CCL-185) were cultured as a monolayer in DMEM (Gibco, 10566016), supplemented with 10% v/v FBS (Thermo Fisher), 100 U ml-1 penicillin (Thermo Fisher), and 100 μg ml-1 streptomycin (Thermo Fisher) on 15 cm Nunclon Delta dishes (Thermo Fisher). ..
Cell Culture:Article Title: Multiprobe Photoproximity Labeling of the EGFR Interactome in Glioblastoma Using Red-Light
Article Snippet: Duolink® Proximity Ligation Assay kits (Duolink® In Situ PLA® Probe Anti-Rabbit PLUS, DUO92002; Duolink® In Situ PLA® Probe Anti-Mouse MINUS, DUO92004; Duolink® In Situ Detection Reagents Orange, DUO92007) were purchased from EMD Millipore. .. Structure of probes used in this study HN NH S O N H O NH2 H H O N H HN NH S O OH H H O N H HN NH S O H NO O O O H H CF3 NN O N H HN NH S O N H O O O H H O N3 O H N NH2NH O HN NH S O H H Dz-Bt Az-Bt Hy-BtPhenol-Bt An-Bt Biotin-Phenol (Biotinyl tyramide) was purchased from SIGMA (SML2135); Biotin LC hydrazide was purchased from Santa Cruz Biotechnology (sc-257155); Biotin-aniline was purchased from SIGMA (SML2946); Biotin-PEG3-Ph(4-N3) was purchased from Iris Biotech GMBH (PEG6795); Biotin-PEG3Diazrine was prepared accoriding to the literature.1 Recombinant protein used in this study Carbonic anhydrase (CA) was purchased from Mp Biomedicals Inc (ICN15387925), Bovine serum albumin (BSA) was purchased from SIGMA (A9647); Recombinant Human Epidermal growth factor (EGF) was purchased from Thermo Fisher scientific (AF-100-15-1mg) Antibodies used in this study Streptavidin HRP-Conjugated Streptavidin , Thermo Fisher scientific, N100 1:1000 in TBST Western Blot Streptavidin IRDye® 800CW Streptavidin Li Cor Inc, 926-32230 1:1000 in TBST Western Blot Streptavidin IRDye® 680LT Streptavidin 1:1000 in TBST 34 Li Cor Inc, 926-68031 Western Blot Streptavidin DyLightTM 550, Thermo Fisher scientific, 84542 1:1000 in DPBS Flow Cyctometry Goat anti-Rabbit IRDye® 800CW, 926-32211 1:10000 in TBSTWestern Blot Cetuximab (anti-EGFR) Selleck Chemicals LLC, A2000 Anti-EGFR Cell Signaling Technology, 4267S 1:500 in TBSTWestern Blot Human IgG Thermo Fisher scientific, 0931431 Anti-EGFR BD PharmingenTM, 555330 1:200 in Proximity Ligation Assay Anti-NEO1 Thermo Fisher scientific, 20246-1-AP 1:200 in Proximity Ligation Assay Goat anti-Human-IgG 647 Thermo Fisher scientific, A21445 1:1000 in DPBSFlow Cyctometry Anti-β-actin Cell Signaling Technology, 3700 1:500 in TBSTWestern Blot Goat anti-Mice IRDye® 680CW, 926-68070 1:10000 in TBSTWestern Blot Cell Culture HEK 293T cells (ATCC CRL-3216) and A549 cells (ATCC CCL-185) were cultured as a monolayer in DMEM (Gibco, 10566016), supplemented with 10% v/v FBS (Thermo Fisher), 100 U ml-1 penicillin (Thermo Fisher), and 100 μg ml-1 streptomycin (Thermo Fisher) on 15 cm Nunclon Delta dishes (Thermo Fisher). ..
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