Immunoprecipitation:Article Title: Breast cancer cell-derived extracellular vesicles promote CD8 + T cell exhaustion via TGF-β type II receptor signaling
Article Snippet: .. The antibodies used for immunoprecipitation (IP), immunoblotting (IB), immunofluorescence (IF), and immunohistochemistry (IHC) were as follows: TβRII (sc-400, Santa Cruz Biotechnology, 1:1000 for IB, 1:50 for IP, 1:100 for IHC, 1:50 for immunogold label, 1:100 for IF), human TβRII (ab184948, Abcam, 1:1000 for IB), CD63 (ab216130, Abcam, 1:2000 for IB), TSG101 (sc-7964, Santa Cruz Biotechnology, 1:1000 for IB), Alix (sc-53540, Santa Cruz Biotechnology, 1:1000 for IB, 1:100 for IF), CD9 (A19027, ABclonal, 1:2000 for IB), CD81(sc-166029, Santa Cruz Biotechnology, 1:1000 for IB), Calnexin (A4846, ABclonal, 1:2000 for IB), Hrs (A1790, ABclonal, 1:2000 for IB), Rab27a (sc-74586, Santa Cruz Biotechnology, 1:1000 for IB), N-cadherin (610920, BD Bioscience, 1:50000 for IB), E-cadherin (610181, BD Bioscience, 1:10000 for IB, 1:100 for IF), SMAD4 (sc-7966, Santa Cruz Biotechnology, 1:1000 for IB), SMAD2-3 (610842, BD Bioscience, 1:2500 for IB, 1:500 for IP, 1:100 for IF), phospho-SMAD2 (#3101, Cell Signaling, 1:5000 for IB, 1:50 for IHC), SMAD3 (A19115, ABclonal, 1:100 for IP, 1:2000 for IB), phospho-SMAD3 (AP0727, ABclonal, 1:2000 for IB), SMAD4 (A19116, ABclonal, 1:2000 for IB), Ub (sc-8017, Santa Cruz Biotechnology, 1:1000 for IB), fibronectin (SAB4500974, Sigma, 1:1000 for IB), SMA (SAB5500002, Sigma, 1:1000 for IB), vimentin (#5741, Cell Signaling, 1:1000 for IB), TCF1/7 (#2203, Cell Signaling, 1:1000 for IB), EOMES (#4540, Cell Signaling, 1:1000 for IB), GZMB (sc-8022, Santa Cruz Biotechnology, 1:1000 for IB), CD8a (ab22378, Abcam, 1:400 for IHC), K48-linkage specific polyubiquitin (#8081 S, Cell Signaling, 1:1000 for IB), K63-linkage specific polyubiquitin (#A18164, ABclonal, 1:1000 for IB), β-actin (#A5441, Sigma, 1:10000 for IB), Phalloidin (#93042, Sigma, 1:1000 for IF), polyclonal HA(Y-11) (sc-805, Santa Cruz Biotechnology, 1:1000 for IB), monoclonal HA(12CAS5, home-made, 1:5000 for IB), polyclonal Myc (A-14) (sc-789, Santa Cruz Biotechnology, 1:1000 for IB), monoclonal Myc (9E10) (sc-40, Santa Cruz Biotechnology, 1:1000 for IB), Flag (M2, Sigma, 1:10000 for IB, 1:200 for IF), Protein A–HRP (Sigma-Aldrich GENA9120, 1:10000 for IB), HRP-conjugated secondary antibodies to mouse (NA931) or rabbit (NA934) (both from Amersham Biosciences, 1:10000 for IB), AlexaFluor488-labeled secondary antibody to rabbit (Molecular Probes R37116, 1:300 for IF) or AlexaFluor593-labeled antibody to mouse (Molecular Probes R3712, 1:300 for IF). ..
Western Blot:Article Title: Breast cancer cell-derived extracellular vesicles promote CD8 + T cell exhaustion via TGF-β type II receptor signaling
Article Snippet: .. The antibodies used for immunoprecipitation (IP), immunoblotting (IB), immunofluorescence (IF), and immunohistochemistry (IHC) were as follows: TβRII (sc-400, Santa Cruz Biotechnology, 1:1000 for IB, 1:50 for IP, 1:100 for IHC, 1:50 for immunogold label, 1:100 for IF), human TβRII (ab184948, Abcam, 1:1000 for IB), CD63 (ab216130, Abcam, 1:2000 for IB), TSG101 (sc-7964, Santa Cruz Biotechnology, 1:1000 for IB), Alix (sc-53540, Santa Cruz Biotechnology, 1:1000 for IB, 1:100 for IF), CD9 (A19027, ABclonal, 1:2000 for IB), CD81(sc-166029, Santa Cruz Biotechnology, 1:1000 for IB), Calnexin (A4846, ABclonal, 1:2000 for IB), Hrs (A1790, ABclonal, 1:2000 for IB), Rab27a (sc-74586, Santa Cruz Biotechnology, 1:1000 for IB), N-cadherin (610920, BD Bioscience, 1:50000 for IB), E-cadherin (610181, BD Bioscience, 1:10000 for IB, 1:100 for IF), SMAD4 (sc-7966, Santa Cruz Biotechnology, 1:1000 for IB), SMAD2-3 (610842, BD Bioscience, 1:2500 for IB, 1:500 for IP, 1:100 for IF), phospho-SMAD2 (#3101, Cell Signaling, 1:5000 for IB, 1:50 for IHC), SMAD3 (A19115, ABclonal, 1:100 for IP, 1:2000 for IB), phospho-SMAD3 (AP0727, ABclonal, 1:2000 for IB), SMAD4 (A19116, ABclonal, 1:2000 for IB), Ub (sc-8017, Santa Cruz Biotechnology, 1:1000 for IB), fibronectin (SAB4500974, Sigma, 1:1000 for IB), SMA (SAB5500002, Sigma, 1:1000 for IB), vimentin (#5741, Cell Signaling, 1:1000 for IB), TCF1/7 (#2203, Cell Signaling, 1:1000 for IB), EOMES (#4540, Cell Signaling, 1:1000 for IB), GZMB (sc-8022, Santa Cruz Biotechnology, 1:1000 for IB), CD8a (ab22378, Abcam, 1:400 for IHC), K48-linkage specific polyubiquitin (#8081 S, Cell Signaling, 1:1000 for IB), K63-linkage specific polyubiquitin (#A18164, ABclonal, 1:1000 for IB), β-actin (#A5441, Sigma, 1:10000 for IB), Phalloidin (#93042, Sigma, 1:1000 for IF), polyclonal HA(Y-11) (sc-805, Santa Cruz Biotechnology, 1:1000 for IB), monoclonal HA(12CAS5, home-made, 1:5000 for IB), polyclonal Myc (A-14) (sc-789, Santa Cruz Biotechnology, 1:1000 for IB), monoclonal Myc (9E10) (sc-40, Santa Cruz Biotechnology, 1:1000 for IB), Flag (M2, Sigma, 1:10000 for IB, 1:200 for IF), Protein A–HRP (Sigma-Aldrich GENA9120, 1:10000 for IB), HRP-conjugated secondary antibodies to mouse (NA931) or rabbit (NA934) (both from Amersham Biosciences, 1:10000 for IB), AlexaFluor488-labeled secondary antibody to rabbit (Molecular Probes R37116, 1:300 for IF) or AlexaFluor593-labeled antibody to mouse (Molecular Probes R3712, 1:300 for IF). ..
Immunofluorescence:Article Title: Breast cancer cell-derived extracellular vesicles promote CD8 + T cell exhaustion via TGF-β type II receptor signaling
Article Snippet: .. The antibodies used for immunoprecipitation (IP), immunoblotting (IB), immunofluorescence (IF), and immunohistochemistry (IHC) were as follows: TβRII (sc-400, Santa Cruz Biotechnology, 1:1000 for IB, 1:50 for IP, 1:100 for IHC, 1:50 for immunogold label, 1:100 for IF), human TβRII (ab184948, Abcam, 1:1000 for IB), CD63 (ab216130, Abcam, 1:2000 for IB), TSG101 (sc-7964, Santa Cruz Biotechnology, 1:1000 for IB), Alix (sc-53540, Santa Cruz Biotechnology, 1:1000 for IB, 1:100 for IF), CD9 (A19027, ABclonal, 1:2000 for IB), CD81(sc-166029, Santa Cruz Biotechnology, 1:1000 for IB), Calnexin (A4846, ABclonal, 1:2000 for IB), Hrs (A1790, ABclonal, 1:2000 for IB), Rab27a (sc-74586, Santa Cruz Biotechnology, 1:1000 for IB), N-cadherin (610920, BD Bioscience, 1:50000 for IB), E-cadherin (610181, BD Bioscience, 1:10000 for IB, 1:100 for IF), SMAD4 (sc-7966, Santa Cruz Biotechnology, 1:1000 for IB), SMAD2-3 (610842, BD Bioscience, 1:2500 for IB, 1:500 for IP, 1:100 for IF), phospho-SMAD2 (#3101, Cell Signaling, 1:5000 for IB, 1:50 for IHC), SMAD3 (A19115, ABclonal, 1:100 for IP, 1:2000 for IB), phospho-SMAD3 (AP0727, ABclonal, 1:2000 for IB), SMAD4 (A19116, ABclonal, 1:2000 for IB), Ub (sc-8017, Santa Cruz Biotechnology, 1:1000 for IB), fibronectin (SAB4500974, Sigma, 1:1000 for IB), SMA (SAB5500002, Sigma, 1:1000 for IB), vimentin (#5741, Cell Signaling, 1:1000 for IB), TCF1/7 (#2203, Cell Signaling, 1:1000 for IB), EOMES (#4540, Cell Signaling, 1:1000 for IB), GZMB (sc-8022, Santa Cruz Biotechnology, 1:1000 for IB), CD8a (ab22378, Abcam, 1:400 for IHC), K48-linkage specific polyubiquitin (#8081 S, Cell Signaling, 1:1000 for IB), K63-linkage specific polyubiquitin (#A18164, ABclonal, 1:1000 for IB), β-actin (#A5441, Sigma, 1:10000 for IB), Phalloidin (#93042, Sigma, 1:1000 for IF), polyclonal HA(Y-11) (sc-805, Santa Cruz Biotechnology, 1:1000 for IB), monoclonal HA(12CAS5, home-made, 1:5000 for IB), polyclonal Myc (A-14) (sc-789, Santa Cruz Biotechnology, 1:1000 for IB), monoclonal Myc (9E10) (sc-40, Santa Cruz Biotechnology, 1:1000 for IB), Flag (M2, Sigma, 1:10000 for IB, 1:200 for IF), Protein A–HRP (Sigma-Aldrich GENA9120, 1:10000 for IB), HRP-conjugated secondary antibodies to mouse (NA931) or rabbit (NA934) (both from Amersham Biosciences, 1:10000 for IB), AlexaFluor488-labeled secondary antibody to rabbit (Molecular Probes R37116, 1:300 for IF) or AlexaFluor593-labeled antibody to mouse (Molecular Probes R3712, 1:300 for IF). ..
Immunohistochemistry:Article Title: Breast cancer cell-derived extracellular vesicles promote CD8 + T cell exhaustion via TGF-β type II receptor signaling
Article Snippet: .. The antibodies used for immunoprecipitation (IP), immunoblotting (IB), immunofluorescence (IF), and immunohistochemistry (IHC) were as follows: TβRII (sc-400, Santa Cruz Biotechnology, 1:1000 for IB, 1:50 for IP, 1:100 for IHC, 1:50 for immunogold label, 1:100 for IF), human TβRII (ab184948, Abcam, 1:1000 for IB), CD63 (ab216130, Abcam, 1:2000 for IB), TSG101 (sc-7964, Santa Cruz Biotechnology, 1:1000 for IB), Alix (sc-53540, Santa Cruz Biotechnology, 1:1000 for IB, 1:100 for IF), CD9 (A19027, ABclonal, 1:2000 for IB), CD81(sc-166029, Santa Cruz Biotechnology, 1:1000 for IB), Calnexin (A4846, ABclonal, 1:2000 for IB), Hrs (A1790, ABclonal, 1:2000 for IB), Rab27a (sc-74586, Santa Cruz Biotechnology, 1:1000 for IB), N-cadherin (610920, BD Bioscience, 1:50000 for IB), E-cadherin (610181, BD Bioscience, 1:10000 for IB, 1:100 for IF), SMAD4 (sc-7966, Santa Cruz Biotechnology, 1:1000 for IB), SMAD2-3 (610842, BD Bioscience, 1:2500 for IB, 1:500 for IP, 1:100 for IF), phospho-SMAD2 (#3101, Cell Signaling, 1:5000 for IB, 1:50 for IHC), SMAD3 (A19115, ABclonal, 1:100 for IP, 1:2000 for IB), phospho-SMAD3 (AP0727, ABclonal, 1:2000 for IB), SMAD4 (A19116, ABclonal, 1:2000 for IB), Ub (sc-8017, Santa Cruz Biotechnology, 1:1000 for IB), fibronectin (SAB4500974, Sigma, 1:1000 for IB), SMA (SAB5500002, Sigma, 1:1000 for IB), vimentin (#5741, Cell Signaling, 1:1000 for IB), TCF1/7 (#2203, Cell Signaling, 1:1000 for IB), EOMES (#4540, Cell Signaling, 1:1000 for IB), GZMB (sc-8022, Santa Cruz Biotechnology, 1:1000 for IB), CD8a (ab22378, Abcam, 1:400 for IHC), K48-linkage specific polyubiquitin (#8081 S, Cell Signaling, 1:1000 for IB), K63-linkage specific polyubiquitin (#A18164, ABclonal, 1:1000 for IB), β-actin (#A5441, Sigma, 1:10000 for IB), Phalloidin (#93042, Sigma, 1:1000 for IF), polyclonal HA(Y-11) (sc-805, Santa Cruz Biotechnology, 1:1000 for IB), monoclonal HA(12CAS5, home-made, 1:5000 for IB), polyclonal Myc (A-14) (sc-789, Santa Cruz Biotechnology, 1:1000 for IB), monoclonal Myc (9E10) (sc-40, Santa Cruz Biotechnology, 1:1000 for IB), Flag (M2, Sigma, 1:10000 for IB, 1:200 for IF), Protein A–HRP (Sigma-Aldrich GENA9120, 1:10000 for IB), HRP-conjugated secondary antibodies to mouse (NA931) or rabbit (NA934) (both from Amersham Biosciences, 1:10000 for IB), AlexaFluor488-labeled secondary antibody to rabbit (Molecular Probes R37116, 1:300 for IF) or AlexaFluor593-labeled antibody to mouse (Molecular Probes R3712, 1:300 for IF). ..
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