polyclonal rabbit cdc25c antibody  (Santa Cruz Biotechnology)

Journal: Retrovirology

Article Title: Human T-lymphotropic virus type-1 p30 alters cell cycle G2 regulation of T lymphocytes to enhance cell survival

doi: 10.1186/1742-4690-4-49

Figure Lengend Snippet: p30 enhances phosphorylation of Cdc25C at S-216 . A) 40 ug of nuclear (N) or cytosolic (C) fraction prepared from either p30 expressing or Mock Jurkat T-cells were loaded on 10% SDS gel and western blot analysis was performed with anti-HA to confirm p30 expression and Histone H1 western to confirm nuclear and cytosolic fractionation. B) Western blot of nuclear (N) and cytosolic (C) extracts prepared from p30 or mock Jurkat T-cells probed with anti-Cdc2 and phosphospecific Cdc2 antibody. C) Western blot of nuclear (N) and cytosolic (C) extracts prepared from p30 or mock Jurkat T-cells, probed with anti-Cdc25C or phosphospecific anti-Cdc25C. D) Western Blot analysis of nuclear or cytosolic extracts from p30 or mock Jurkat T-cells, probed with anti-Chk1 or monoclonal phosphospecific Chk1 (S-345).

Article Snippet: Immunodetection was performed using the following antibodies: mouse anti-HA monoclonal antibody clone 16B-12 (1:1000, Covance Research Products, Princeton, NJ), mouse monoclonal anti-PARP clone C-2-10 (1:1000, Oncogene Research Products, Boston, MA). mouse monoclonal anti-β-actin clone AC-74 (1:4000, Sigma), rabbit polyclonal anti-Cdc2 # 9112 (1:1000, Cell Signaling, Beverly, MA), rabbit polyclonal anti-PLK1pT210 # 600-401-466 (1: 1000, Rockland, Gilbertsville, PA), polyclonal rabbit anti-Cdc2 T-161 #9114 (1:1000, Cell Signaling), polyclonal rabbit anti Cdc2 Y-15 #9111(1:1000, Cell Signaling), polyclonal rabbit anti-Cdc25C #9522 (1:1000, Cell Signaling), polyclonal rabbit anti-Cdc25C S-216 # 9528(1:1000, Cell Signaling), rabbit polyclonal anti-Cdc25C pS-198 # 9529 (1: 1000, Cell Signaling), polyclonal rabbit anti-Chk1 #2345 (1:1000, Cell Signaling), polyclonal rabbit anti-Chk1 S-345 #2341 (1: 1000, Cell Signaling), rabbit polyclonal anti-PLK1 (1: 1000) (Abcam, Cambridge, MA), mouse monoclonal anti-Histone H1, clone AE-4 (1: 1000, Upstate).

Techniques: Expressing, SDS-Gel, Western Blot, Fractionation

Journal: Retrovirology

Article Title: Human T-lymphotropic virus type-1 p30 alters cell cycle G2 regulation of T lymphocytes to enhance cell survival

doi: 10.1186/1742-4690-4-49

Figure Lengend Snippet: p30 enhances phosphorylation of Cdc25C at S-216 . A) 40 ug of nuclear (N) or cytosolic (C) fraction prepared from either p30 expressing or Mock Jurkat T-cells were loaded on 10% SDS gel and western blot analysis was performed with anti-HA to confirm p30 expression and Histone H1 western to confirm nuclear and cytosolic fractionation. B) Western blot of nuclear (N) and cytosolic (C) extracts prepared from p30 or mock Jurkat T-cells probed with anti-Cdc2 and phosphospecific Cdc2 antibody. C) Western blot of nuclear (N) and cytosolic (C) extracts prepared from p30 or mock Jurkat T-cells, probed with anti-Cdc25C or phosphospecific anti-Cdc25C. D) Western Blot analysis of nuclear or cytosolic extracts from p30 or mock Jurkat T-cells, probed with anti-Chk1 or monoclonal phosphospecific Chk1 (S-345).

Article Snippet: Immunodetection was performed using the following antibodies: mouse anti-HA monoclonal antibody clone 16B-12 (1:1000, Covance Research Products, Princeton, NJ), mouse monoclonal anti-PARP clone C-2-10 (1:1000, Oncogene Research Products, Boston, MA). mouse monoclonal anti-β-actin clone AC-74 (1:4000, Sigma), rabbit polyclonal anti-Cdc2 # 9112 (1:1000, Cell Signaling, Beverly, MA), rabbit polyclonal anti-PLK1pT210 # 600-401-466 (1: 1000, Rockland, Gilbertsville, PA), polyclonal rabbit anti-Cdc2 T-161 #9114 (1:1000, Cell Signaling), polyclonal rabbit anti Cdc2 Y-15 #9111(1:1000, Cell Signaling), polyclonal rabbit anti-Cdc25C #9522 (1:1000, Cell Signaling), polyclonal rabbit anti-Cdc25C S-216 # 9528(1:1000, Cell Signaling), rabbit polyclonal anti-Cdc25C pS-198 # 9529 (1: 1000, Cell Signaling), polyclonal rabbit anti-Chk1 #2345 (1:1000, Cell Signaling), polyclonal rabbit anti-Chk1 S-345 #2341 (1: 1000, Cell Signaling), rabbit polyclonal anti-PLK1 (1: 1000) (Abcam, Cambridge, MA), mouse monoclonal anti-Histone H1, clone AE-4 (1: 1000, Upstate).

Techniques: Expressing, SDS-Gel, Western Blot, Fractionation

Journal: Retrovirology

Article Title: Human T-lymphotropic virus type-1 p30 alters cell cycle G2 regulation of T lymphocytes to enhance cell survival

doi: 10.1186/1742-4690-4-49

Figure Lengend Snippet: p30 enhances phosphorylation of Cdc25C at S-216 . A) 40 ug of nuclear (N) or cytosolic (C) fraction prepared from either p30 expressing or Mock Jurkat T-cells were loaded on 10% SDS gel and western blot analysis was performed with anti-HA to confirm p30 expression and Histone H1 western to confirm nuclear and cytosolic fractionation. B) Western blot of nuclear (N) and cytosolic (C) extracts prepared from p30 or mock Jurkat T-cells probed with anti-Cdc2 and phosphospecific Cdc2 antibody. C) Western blot of nuclear (N) and cytosolic (C) extracts prepared from p30 or mock Jurkat T-cells, probed with anti-Cdc25C or phosphospecific anti-Cdc25C. D) Western Blot analysis of nuclear or cytosolic extracts from p30 or mock Jurkat T-cells, probed with anti-Chk1 or monoclonal phosphospecific Chk1 (S-345).

Article Snippet: Immunodetection was performed using the following antibodies: mouse anti-HA monoclonal antibody clone 16B-12 (1:1000, Covance Research Products, Princeton, NJ), mouse monoclonal anti-PARP clone C-2-10 (1:1000, Oncogene Research Products, Boston, MA). mouse monoclonal anti-β-actin clone AC-74 (1:4000, Sigma), rabbit polyclonal anti-Cdc2 # 9112 (1:1000, Cell Signaling, Beverly, MA), rabbit polyclonal anti-PLK1pT210 # 600-401-466 (1: 1000, Rockland, Gilbertsville, PA), polyclonal rabbit anti-Cdc2 T-161 #9114 (1:1000, Cell Signaling), polyclonal rabbit anti Cdc2 Y-15 #9111(1:1000, Cell Signaling), polyclonal rabbit anti-Cdc25C #9522 (1:1000, Cell Signaling), polyclonal rabbit anti-Cdc25C S-216 # 9528(1:1000, Cell Signaling), rabbit polyclonal anti-Cdc25C pS-198 # 9529 (1: 1000, Cell Signaling), polyclonal rabbit anti-Chk1 #2345 (1:1000, Cell Signaling), polyclonal rabbit anti-Chk1 S-345 #2341 (1: 1000, Cell Signaling), rabbit polyclonal anti-PLK1 (1: 1000) (Abcam, Cambridge, MA), mouse monoclonal anti-Histone H1, clone AE-4 (1: 1000, Upstate).

Techniques: Expressing, SDS-Gel, Western Blot, Fractionation

Journal: Molecular Cancer

Article Title: Apigenin inhibits pancreatic cancer cell proliferation through G2/M cell cycle arrest

doi: 10.1186/1476-4598-5-76

Figure Lengend Snippet: Effects of apigenin on proteins involved in the G2/M cell cycle phase transition . Expression of both cyclin A and cyclin B were both decreased in apigenin-treated cells compared to controls. While the expression of the cdc2 protein was unchanged, levels of phosphorylated cdc2 were decreased in treated cells. Expression of cdc25A and cdc25C were also decreased in treated cells compared to controls.

Article Snippet: The monoclonal mouse cyclin A, B, cdc25A, and GAPDH antibodies, and the polyclonal rabbit cdc25C antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Sublimation, Expressing

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Ethanol Extract of Root of Prunus persica Inhibited the Growth of Liver Cancer Cell HepG2 by Inducing Cell Cycle Arrest and Migration Suppression

doi: 10.1155/2017/8231936

Figure Lengend Snippet: TSG treatment induced G2/M phase arrest of HepG2 cells. (a) Flow cytometry analysis of DNA content of HepG2 cells subjected to indicated treatment. ((b), (c)) Statistical analysis of cell subpopulation ratios for each phase of cell cycle. ∗ p < 0.05. ∗∗ p < 0.01. ∗∗∗ p < 0.001. (d) Western blotting analysis to detect the expression of CDK1 and Cdc25c.

Article Snippet: Antibodies used in this study included rabbit anti-Cdc25c polyclonal antibody (Cat# D154112, Sangon Bio-tech, China), rabbit anti-CDK1 polyclonal antibody (Cat# D160158, Sangon Bio-tech, China), rabbit anti-MMP9 polyclonal antibody (Cat# 10375-2-AP, Proteintech, USA), rabbit anti-MMP3 polyclonal antibody (Cat# 17873-1-AP, Proteintech, USA), mouse anti- β -Actin monoclonal antibody (Cat# TA-09, ZSGB-Bio, China), HRP-conjugated goat anti-mouse polyclonal antibody (Cat# ZB2305, ZSGB-Bio, China), and HRP-conjugated goat anti-rabbit polyclonal antibody (Cat# ZB2301, ZSGB-Bio, China).

Techniques: Flow Cytometry, Western Blot, Expressing

Journal: Molecular Cancer

Article Title: RNA demethylase ALKBH5 prevents pancreatic cancer progression by posttranscriptional activation of PER1 in an m6A-YTHDF2-dependent manner

doi: 10.1186/s12943-020-01158-w

Figure Lengend Snippet: ALKBH5 induces posttranscriptional activation of PER1 to inhibit PC progression in an m6A-YTHDF2-dependent manner. ALKBH5 loss that abrogates mRNA demethylation leads to an upregulation of PER1 m6A levels in PC cells. Then, degradation of PER1 mRNA occurs on the basis of the m6A-YTHDF2 interaction. PER1 downregulation results in the inhibition of ATM phosphorylation and inactivation of the ATM-CHK2-P53/CDC25C pathways. G2/M arrest in PC cells was suppressed as a result. The PER1-related P53 downregulation and P53-dependent transcriptional inactivation of ALKBH5 indicates a feedback loop underlying the progression of PC

Article Snippet: The primary antibodies were rabbit monoclonal anti-ALKBH5 (ab195377, Abcam), mouse monoclonal anti-PER1 (sc-398,890, Santa Cruz), mouse monoclonal anti-p-ATM (Ser-1981; sc-47,739, Santa Cruz), rabbit monoclonal anti-p-CHK2 (Thr-68; ab32148, Abcam), rabbit polyclonal anti-p-CDC25C (Ser216; ab47322, Abcam), rabbit monoclonal anti-p-P53 (Ser-15; ab1431, Abcam), mouse monoclonal anti-P21 (sc-71,811, Santa Cruz), mouse monoclonal anti-CYCLIN B1 (ab72, Abcam), rabbit polyclonal anti-p-CDK1 (Tyr15; ab47594), mouse monoclonal anti-CDK1 (A17, Abcam) and rabbit polyclonal anti-β-ACTIN (ab8227, Abcam).

Techniques: Activation Assay, Inhibition

Journal: Molecules and Cells

Article Title: Actin Dysfunction Induces Cell Cycle Delay at G2/M with Sustained ERK and RSK Activation in IMR-90 Normal Human Fibroblasts

doi: 10.14348/molcells.2018.2266

Figure Lengend Snippet: As denoted in , IMR-90 cells were synchronized with 2 mM double thymidine arrest, incubated with 5 μM cytochalasin D or the solvent DMSO as a control at 5.5–6 h after the second release, and collected at each indicated time point after the second release. Cell lysates were resolved by 8% SDS-PAGE and blotted. Blots were probed with (A) p-ERK1/2 and p-RSK1 (Ser 380) and re-probed with anti-ERK1/2 and anti-RSK1 to observe the total amount of each protein, (B) p-ERK1/2 and p-Cdc25C (Ser 216), and re-probed with anti-ERK1/2 and anti-Cdc25C. (A, B) Cell cycle progression at G2/M was monitored by detecting p-Cdc2 (Tyr 15) followed by re-probing with anti-Cdc2 to detect the total amount of Cdc2. (C) The same samples from (A) and (B) were blotted with p-Wee1 (Ser 642) and re-probed with anti-Wee1. Each blot was re-probed with anti-actin as a loading control.

Article Snippet: Rabbit polyclonal p-Cdc2 (Tyr 15) (ab47594, Abcam, USA), mouse monoclonal anti-cyclin A (4656, Cell Signaling Technology, USA), rabbit polyclonal p-Cdc25C (Ser 216) (9528, Cell Signaling Technology), rabbit monoclonal anti-Cdc25C (ab32444, Abcam), rabbit polyclonal anti-β-actin (4967, Cell Signaling Technology), rabbit monoclonal p-Wee1 (Ser 642) (4910, Cell Signaling Technology), rabbit polyclonal anti-Wee1 (4936, Cell Signaling Technology), rabbit monoclonal anti-FLAG (F7425, Sigma), rabbit polyclonal p-ERK1/2 (Thr 202/Tyr 204) (9101, Cell Signaling), rabbit polyclonal anti-ERK1/2 (9102, Cell Signaling), rabbit monoclonal p-RSK1 (Ser 380) (ab32203, Abcam), rabbit polyclonal anti-RSK1 (9333, Cell Signaling), rabbit monoclonal histone H3 (4499, Cell Signaling Technology), and mouse monoclonal p-H3 (Ser 10) (9706, Cell Signaling Technology) were used in this study.

Techniques: Incubation, SDS Page

Journal: Molecules and Cells

Article Title: Actin Dysfunction Induces Cell Cycle Delay at G2/M with Sustained ERK and RSK Activation in IMR-90 Normal Human Fibroblasts

doi: 10.14348/molcells.2018.2266

Figure Lengend Snippet: (A) The layout shows how the experiment was performed. IMR-90 cells were synchronized with 2 mM thymidine and transfected with DN-RSK or empty vector as described in the Materials and Methods. Cells were treated with 5 μM CD or only DMSO as a control at 5.5–6 h after the second release and collected at each indicated time point after the second release. Each cell lysate was resolved by 8% SDS-PAGE and blotted with (B) anti-FLAG and anti-actin, (C) p-ERK1/2 and p-Cdc2 (Tyr 15), (D) p-Cdc25C (Ser 216), and p-Wee1 (Ser 642). Each membrane was reprobed with anti-actin as a loading control.

Article Snippet: Rabbit polyclonal p-Cdc2 (Tyr 15) (ab47594, Abcam, USA), mouse monoclonal anti-cyclin A (4656, Cell Signaling Technology, USA), rabbit polyclonal p-Cdc25C (Ser 216) (9528, Cell Signaling Technology), rabbit monoclonal anti-Cdc25C (ab32444, Abcam), rabbit polyclonal anti-β-actin (4967, Cell Signaling Technology), rabbit monoclonal p-Wee1 (Ser 642) (4910, Cell Signaling Technology), rabbit polyclonal anti-Wee1 (4936, Cell Signaling Technology), rabbit monoclonal anti-FLAG (F7425, Sigma), rabbit polyclonal p-ERK1/2 (Thr 202/Tyr 204) (9101, Cell Signaling), rabbit polyclonal anti-ERK1/2 (9102, Cell Signaling), rabbit monoclonal p-RSK1 (Ser 380) (ab32203, Abcam), rabbit polyclonal anti-RSK1 (9333, Cell Signaling), rabbit monoclonal histone H3 (4499, Cell Signaling Technology), and mouse monoclonal p-H3 (Ser 10) (9706, Cell Signaling Technology) were used in this study.

Techniques: Transfection, Plasmid Preparation, SDS Page