Blocking Assay:Article Title: High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells
Article Snippet: .. After 1 h blocking with 5% milk in TBST at room temperature (RT) for 1 h, membranes were incubated overnight at 4 °C with the following primary antibodies: CCND1 (Santa Cruz Biotechnology, sc-20044, 1:500), CCND3 (Santa Cruz Biotechnology, sc-6283, 1:500), CCNE2 (Santa Cruz Biotechnology, sc-248, 1:500), CCNA (Santa Cruz Biotechnology, sc-271682, 1:500), CCNA (Santa Cruz Biotechnology, sc-596, 1:500), CCNB1 (Santa Cruz Biotechnology, sc-245, 1:500), GAPDH (Santa Cruz Biotechnology, sc-51907, 1:500), CDK1 pT161 (Cell Signalling Technology, 9114, 1:1000), CDK1 pY15 (Cell Signalling Technology, 9111, 1:1000), Alpha Tubulin (Abcam, ab52866, 1:5000), PLK1(Millipore, 05-844, 1:1000), CDK2 (Santa Cruz Biotechnology, sc-163, 1:500), CDK1 (Abcam, ab32094, 1:1000), CDC20 (Santa Cruz Biotechnology, sc-13162, 1:500), UBR7 (Thermo Fisher Scientific, A304-130A, 1:1000), UBE2S (Cell Signalling Technology, 11878, 1:1000), CDKN1B (Dako, M7203, 1:1000), CDKN1B pS10 (Santa Cruz Biotechnology, sc-12939, 1:500), NCL (Abcam, ab22758, 1:2000), NCL pT76 (Abcam, ab168363, 1:1000), CCNB1 (Santa Cruz Biotechnology, sc-245, 1:500), SGO2 (Thermo Fisher Scientific, A301-261A, 1:1000), GTSE1 (Abnova, H00051512-B01P, 1:1000), MELK (Thermo Fisher Scientific, A303-136A, 1:1000), FAM83D (Custom antibody from Erich Nigg, 1:500), ANAPC4 (Thermo Fisher Scientific, A301-175A, 1:1000), Phospho-CDK substrate motif [(K/H)pSP] (Cell Signalling Technology, 9477, 1:1000), JUNB (Proteintech, 10486-1-AP, 1:1000), CHAF1B (Thermo Fisher Scientific, A301-085A, 1:2000), Histone H3 pS10 (Cell Signalling Technology, 3377, 1:1000), Actin (Sigma-Aldrich, A5441, 1:5000). .. After washed with TBST for three times, membranes were incubated for 1 h at RT with the following secondary antibodies: IRDye 680RD Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-68073, 1:10000), IRDye 800CW Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-32213, 1:10000), IRDye 800CW Donkey anti-Mouse IgG (H + L) (LI-COR Biosciences, 926-32212, 1:10000), IRDye 680RD Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-68073, 1:10000), anti-mouse HRP (Cell Signalling Technology, 7076, 1:2000), anti-rabbit HRP (Cell Signalling Technology, 7074, 1:2000).
Article Title: High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells
Article Snippet: .. After 1 h blocking with 5% milk in TBST at room temperature (RT) for 1 h, membranes were incubated overnight at 4°C with the following primary antibodies: CCND1 (Santa Cruz Biotechnology, sc-2004), CCND3 (Santa Cruz Biotechnology, sc-6283), CCNE2 (Santa Cruz Biotechnology, sc-248), CCNA (Santa Cruz Biotechnology, sc-271682), CCNA (Santa Cruz Biotechnology, sc-596), CCNB1 (Santa Cruz Biotechnology, sc-245), GAPDH (Santa Cruz Biotechnology, sc-51907), CDK1 pT161 (Cell Signalling Technology, 9114), CDK1 pY15 (Cell Signalling Technology, 9111), Alpha Tubulin (Abcam, ab52866), PLK1(Millipore, 05-844), CDK2 (Santa Cruz Biotechnology, sc-163), CDK1 (Abcam, ab32094), CDC20 (Santa Cruz Biotechnology, sc-13162), UBR7 (Thermo Fisher Scientific, A304-130A), UBE2S (Cell Signalling Technology, 11878), CDKN1B (Dako, M7203), CDKN1B pS10 (Santa Cruz Biotechnology, sc-12939), NCL (Abcam, ab22758), NCL pT76 (Abcam, ab168363), CCNB1 (Santa Cruz Biotechnology, sc-245), SGO2 (Thermo Fisher Scientific, A301-261A), GTSE1 (Abnova, H00051512-B01P), MELK (Thermo Fisher Scientific, A303-136A), FAM83D (Custom antibody from Erich Nigg), ANAPC4 (Thermo Fisher Scientific, A301-175A), Phospho-CDK substrate motif [(K/H)pSP] (Cell Signalling Technology, 9477), JUNB (Proteintech, 10486-1-AP), CHAF1B (Thermo Fisher Scientific, A301-085A), Histone H3 pS10 (Cell Signalling Technology, 3377), Actin (Sigma-Aldrich, A5441). .. After washed with TBST for three times, membranes were incubated for 1h at RT with the following secondary antibodies: IRDye 680RD Donkey anti-Rabbit IgG (H+L) (LI-COR Biosciences, 926-68073), IRDye 800CW Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-32213), IRDye 800CW Donkey anti-Mouse IgG (H + L) (LI-COR Biosciences, 926-32212), IRDye 680RD Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-68073), anti-mouse HRP (Cell Signalling Technology, 7076), anti-rabbit HRP (Cell Signalling Technology, 7074).
Article Title: High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells.
Article Snippet: .. After 1 h blocking with 5% milk in TBST at room temperature (RT) for 1 h, membranes were incubated overnight at 4 °C with the following primary antibodies: CCND1 (Santa Cruz Biotechnology, sc-20044, 1:500), CCND3 (Santa Cruz Biotechnology, sc-6283, 1:500), CCNE2 (Santa Cruz Biotechnology, sc-248, 1:500), CCNA (Santa Cruz Biotechnology, sc-271682, 1:500), CCNA (Santa Cruz Biotechnology, sc-596, 1:500), CCNB1 (Santa Cruz Biotechnology, sc-245, 1:500), GAPDH (Santa Cruz Biotechnology, sc-51907, 1:500), CDK1 pT161 (Cell Signalling Technology, 9114, 1:1000), CDK1 pY15 (Cell Signalling Technology, 9111, 1:1000), Alpha Tubulin (Abcam, ab52866, 1:5000), PLK1(Millipore, 05-844, 1:1000), CDK2 (Santa Cruz Biotechnology, sc-163, 1:500), CDK1 (Abcam, ab32094, 1:1000), CDC20 (Santa Cruz Biotechnology, sc-13162, 1:500), UBR7 (Thermo Fisher Scientific, A304-130A, 1:1000), UBE2S (Cell Signalling Technology, 11878, 1:1000), CDKN1B (Dako, M7203, 1:1000), CDKN1B pS10 (Santa Cruz Biotechnology, sc-12939, 1:500), NCL (Abcam, ab22758, 1:2000), NCL pT76 (Abcam, ab168363, 1:1000), CCNB1 (Santa Cruz Biotechnology, sc-245, 1:500), SGO2 (Thermo Fisher Scientific,A301-261A, 1:1000), GTSE1 (Abnova, H00051512-B01P, 1:1000), MELK (Thermo Fisher Scientific, A303-136A, 1:1000), FAM83D (Custom antibody from Erich Nigg, 1:500), ANAPC4 (Thermo Fisher Scientific, A301-175A, 1:1000), Phospho-CDK substrate motif [(K/H) pSP] (Cell Signalling Technology, 9477, 1:1000), JUNB (Proteintech, 10486-1-AP, 1:1000), CHAF1B (Thermo Fisher Scientific, A301-085A, 1:2000), Histone H3 pS10 (Cell Signalling Technology, 3377, 1:1000), Actin (Sigma-Aldrich, A5441, 1:5000). .. Afterwashedwith TBST for three times, membranes were incubated for 1 h at RT with the following secondary antibodies: IRDye 680RDDonkey anti-Rabbit IgG (H + L) (LICOR Biosciences, 926-68073, 1:10000), IRDye 800CW Donkey antiRabbit IgG (H+ L) (LI-COR Biosciences, 926-32213, 1:10000), IRDye 800CW Donkey anti-Mouse IgG (H+ L) (LI-COR Biosciences, 926- 32212, 1:10000), IRDye 680RD Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-68073, 1:10000), anti-mouse HRP (Cell Signalling Technology, 7076, 1:2000), anti-rabbit HRP (Cell Signalling Technology, 7074, 1:2000).
Incubation:Article Title: High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells
Article Snippet: .. After 1 h blocking with 5% milk in TBST at room temperature (RT) for 1 h, membranes were incubated overnight at 4 °C with the following primary antibodies: CCND1 (Santa Cruz Biotechnology, sc-20044, 1:500), CCND3 (Santa Cruz Biotechnology, sc-6283, 1:500), CCNE2 (Santa Cruz Biotechnology, sc-248, 1:500), CCNA (Santa Cruz Biotechnology, sc-271682, 1:500), CCNA (Santa Cruz Biotechnology, sc-596, 1:500), CCNB1 (Santa Cruz Biotechnology, sc-245, 1:500), GAPDH (Santa Cruz Biotechnology, sc-51907, 1:500), CDK1 pT161 (Cell Signalling Technology, 9114, 1:1000), CDK1 pY15 (Cell Signalling Technology, 9111, 1:1000), Alpha Tubulin (Abcam, ab52866, 1:5000), PLK1(Millipore, 05-844, 1:1000), CDK2 (Santa Cruz Biotechnology, sc-163, 1:500), CDK1 (Abcam, ab32094, 1:1000), CDC20 (Santa Cruz Biotechnology, sc-13162, 1:500), UBR7 (Thermo Fisher Scientific, A304-130A, 1:1000), UBE2S (Cell Signalling Technology, 11878, 1:1000), CDKN1B (Dako, M7203, 1:1000), CDKN1B pS10 (Santa Cruz Biotechnology, sc-12939, 1:500), NCL (Abcam, ab22758, 1:2000), NCL pT76 (Abcam, ab168363, 1:1000), CCNB1 (Santa Cruz Biotechnology, sc-245, 1:500), SGO2 (Thermo Fisher Scientific, A301-261A, 1:1000), GTSE1 (Abnova, H00051512-B01P, 1:1000), MELK (Thermo Fisher Scientific, A303-136A, 1:1000), FAM83D (Custom antibody from Erich Nigg, 1:500), ANAPC4 (Thermo Fisher Scientific, A301-175A, 1:1000), Phospho-CDK substrate motif [(K/H)pSP] (Cell Signalling Technology, 9477, 1:1000), JUNB (Proteintech, 10486-1-AP, 1:1000), CHAF1B (Thermo Fisher Scientific, A301-085A, 1:2000), Histone H3 pS10 (Cell Signalling Technology, 3377, 1:1000), Actin (Sigma-Aldrich, A5441, 1:5000). .. After washed with TBST for three times, membranes were incubated for 1 h at RT with the following secondary antibodies: IRDye 680RD Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-68073, 1:10000), IRDye 800CW Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-32213, 1:10000), IRDye 800CW Donkey anti-Mouse IgG (H + L) (LI-COR Biosciences, 926-32212, 1:10000), IRDye 680RD Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-68073, 1:10000), anti-mouse HRP (Cell Signalling Technology, 7076, 1:2000), anti-rabbit HRP (Cell Signalling Technology, 7074, 1:2000).
Article Title: High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells
Article Snippet: .. After 1 h blocking with 5% milk in TBST at room temperature (RT) for 1 h, membranes were incubated overnight at 4°C with the following primary antibodies: CCND1 (Santa Cruz Biotechnology, sc-2004), CCND3 (Santa Cruz Biotechnology, sc-6283), CCNE2 (Santa Cruz Biotechnology, sc-248), CCNA (Santa Cruz Biotechnology, sc-271682), CCNA (Santa Cruz Biotechnology, sc-596), CCNB1 (Santa Cruz Biotechnology, sc-245), GAPDH (Santa Cruz Biotechnology, sc-51907), CDK1 pT161 (Cell Signalling Technology, 9114), CDK1 pY15 (Cell Signalling Technology, 9111), Alpha Tubulin (Abcam, ab52866), PLK1(Millipore, 05-844), CDK2 (Santa Cruz Biotechnology, sc-163), CDK1 (Abcam, ab32094), CDC20 (Santa Cruz Biotechnology, sc-13162), UBR7 (Thermo Fisher Scientific, A304-130A), UBE2S (Cell Signalling Technology, 11878), CDKN1B (Dako, M7203), CDKN1B pS10 (Santa Cruz Biotechnology, sc-12939), NCL (Abcam, ab22758), NCL pT76 (Abcam, ab168363), CCNB1 (Santa Cruz Biotechnology, sc-245), SGO2 (Thermo Fisher Scientific, A301-261A), GTSE1 (Abnova, H00051512-B01P), MELK (Thermo Fisher Scientific, A303-136A), FAM83D (Custom antibody from Erich Nigg), ANAPC4 (Thermo Fisher Scientific, A301-175A), Phospho-CDK substrate motif [(K/H)pSP] (Cell Signalling Technology, 9477), JUNB (Proteintech, 10486-1-AP), CHAF1B (Thermo Fisher Scientific, A301-085A), Histone H3 pS10 (Cell Signalling Technology, 3377), Actin (Sigma-Aldrich, A5441). .. After washed with TBST for three times, membranes were incubated for 1h at RT with the following secondary antibodies: IRDye 680RD Donkey anti-Rabbit IgG (H+L) (LI-COR Biosciences, 926-68073), IRDye 800CW Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-32213), IRDye 800CW Donkey anti-Mouse IgG (H + L) (LI-COR Biosciences, 926-32212), IRDye 680RD Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-68073), anti-mouse HRP (Cell Signalling Technology, 7076), anti-rabbit HRP (Cell Signalling Technology, 7074).
Article Title: High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells.
Article Snippet: .. After 1 h blocking with 5% milk in TBST at room temperature (RT) for 1 h, membranes were incubated overnight at 4 °C with the following primary antibodies: CCND1 (Santa Cruz Biotechnology, sc-20044, 1:500), CCND3 (Santa Cruz Biotechnology, sc-6283, 1:500), CCNE2 (Santa Cruz Biotechnology, sc-248, 1:500), CCNA (Santa Cruz Biotechnology, sc-271682, 1:500), CCNA (Santa Cruz Biotechnology, sc-596, 1:500), CCNB1 (Santa Cruz Biotechnology, sc-245, 1:500), GAPDH (Santa Cruz Biotechnology, sc-51907, 1:500), CDK1 pT161 (Cell Signalling Technology, 9114, 1:1000), CDK1 pY15 (Cell Signalling Technology, 9111, 1:1000), Alpha Tubulin (Abcam, ab52866, 1:5000), PLK1(Millipore, 05-844, 1:1000), CDK2 (Santa Cruz Biotechnology, sc-163, 1:500), CDK1 (Abcam, ab32094, 1:1000), CDC20 (Santa Cruz Biotechnology, sc-13162, 1:500), UBR7 (Thermo Fisher Scientific, A304-130A, 1:1000), UBE2S (Cell Signalling Technology, 11878, 1:1000), CDKN1B (Dako, M7203, 1:1000), CDKN1B pS10 (Santa Cruz Biotechnology, sc-12939, 1:500), NCL (Abcam, ab22758, 1:2000), NCL pT76 (Abcam, ab168363, 1:1000), CCNB1 (Santa Cruz Biotechnology, sc-245, 1:500), SGO2 (Thermo Fisher Scientific,A301-261A, 1:1000), GTSE1 (Abnova, H00051512-B01P, 1:1000), MELK (Thermo Fisher Scientific, A303-136A, 1:1000), FAM83D (Custom antibody from Erich Nigg, 1:500), ANAPC4 (Thermo Fisher Scientific, A301-175A, 1:1000), Phospho-CDK substrate motif [(K/H) pSP] (Cell Signalling Technology, 9477, 1:1000), JUNB (Proteintech, 10486-1-AP, 1:1000), CHAF1B (Thermo Fisher Scientific, A301-085A, 1:2000), Histone H3 pS10 (Cell Signalling Technology, 3377, 1:1000), Actin (Sigma-Aldrich, A5441, 1:5000). .. Afterwashedwith TBST for three times, membranes were incubated for 1 h at RT with the following secondary antibodies: IRDye 680RDDonkey anti-Rabbit IgG (H + L) (LICOR Biosciences, 926-68073, 1:10000), IRDye 800CW Donkey antiRabbit IgG (H+ L) (LI-COR Biosciences, 926-32213, 1:10000), IRDye 800CW Donkey anti-Mouse IgG (H+ L) (LI-COR Biosciences, 926- 32212, 1:10000), IRDye 680RD Donkey anti-Rabbit IgG (H + L) (LI-COR Biosciences, 926-68073, 1:10000), anti-mouse HRP (Cell Signalling Technology, 7076, 1:2000), anti-rabbit HRP (Cell Signalling Technology, 7074, 1:2000).
Electrophoresis:Article Title: Interaction between ZMIZ2 and AR promotes prostate cancer proliferation in vitro and in vivo.
Article Snippet: .. After electrophoresis, protein was transferred to PVDF and sealed with 5% milk for 1 h. Add following primary antibodies and incubate overnight: ZMIZ2 (Novus Biologicals, LLC, Centennial, CO, USA), AR (Santa Cruz Biotechnology, Dallas, TX, USA), CDK1 (Santa Cruz Biotechnology, Dallas, TX, USA), CCNA2 (Santa Cruz Biotechnology, Dallas, TX, USA), CCNE2 (Santa Cruz Biotechnology, Dallas, TX, USA). .. The protein blots were visualized by employing either a goat anti-rabbit IgG HRPconjugated secondary antibody or a goat anti-mouse IgG HRP-conjugated secondary antibody (purchased from CST, located in Danvers, Massachusetts, USA).
Article Title: Interaction between ZMIZ2 and AR promotes prostate cancer proliferation in vitro and in vivo
Article Snippet: .. After electrophoresis, protein was transferred to PVDF and sealed with 5% milk for 1 h. Add following primary antibodies and incubate overnight: ZMIZ2 (Novus Biologicals, LLC, Centennial, CO, USA), AR (Santa Cruz Biotechnology, Dallas, TX, USA), CDK1 (Santa Cruz Biotechnology, Dallas, TX, USA), CCNA2 (Santa Cruz Biotechnology, Dallas, TX, USA), CCNE2 (Santa Cruz Biotechnology, Dallas, TX, USA). .. The protein blots were visualized by employing either a goat anti-rabbit IgG HRP-conjugated secondary antibody or a goat anti-mouse IgG HRP-conjugated secondary antibody (purchased from CST, located in Danvers, Massachusetts, USA).
Western Blot:Article Title: A patient-derived xenograft mouse platform from epithelioid glioblastoma provides possible druggable screening and translational study
Article Snippet: .. The primary antibodies used in Western blotting and IHC included IDH1 R132H (H09): Dianova, Germany (1:100), Rb: #9313, Cell Signaling Technology, USA (1:1000), p-JNK: #4668, Cell Signaling Technology, USA (1:1000), JNK: #9252, Cell Signaling Technology, USA (1:1000), pRb: #9307, Cell Signaling Technology, USA (1:1000), ERK: SC1647, Santa Cruz Biotechnology, USA (1:500), p-ERK: SC-7383, Santa Cruz Biotechnology, USA (1:500), CCND1: GTX61306, GeneTex, USA (1:1000), CCNE2: 11935-AP, Proteintech (1:1000), CDK4: SC-23896, Santa Cruz Biotechnology, USA (1:1000), CDK6: SC-7961, Santa Cruz Biotechnology, USA (1:1000), β-actin: GT5512, GeneTex, USA (1:10000), BRAF V600E, VE1 (Ventana Medical Systems, automated Ventana Benchmark ULTRA platform). ..
Immunohistochemistry:Article Title: A patient-derived xenograft mouse platform from epithelioid glioblastoma provides possible druggable screening and translational study
Article Snippet: .. The primary antibodies used in Western blotting and IHC included IDH1 R132H (H09): Dianova, Germany (1:100), Rb: #9313, Cell Signaling Technology, USA (1:1000), p-JNK: #4668, Cell Signaling Technology, USA (1:1000), JNK: #9252, Cell Signaling Technology, USA (1:1000), pRb: #9307, Cell Signaling Technology, USA (1:1000), ERK: SC1647, Santa Cruz Biotechnology, USA (1:500), p-ERK: SC-7383, Santa Cruz Biotechnology, USA (1:500), CCND1: GTX61306, GeneTex, USA (1:1000), CCNE2: 11935-AP, Proteintech (1:1000), CDK4: SC-23896, Santa Cruz Biotechnology, USA (1:1000), CDK6: SC-7961, Santa Cruz Biotechnology, USA (1:1000), β-actin: GT5512, GeneTex, USA (1:10000), BRAF V600E, VE1 (Ventana Medical Systems, automated Ventana Benchmark ULTRA platform). ..
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