rabbit anti-human β-actin polyclonal antibody  (Absolute Biotech Inc)

Journal: Cell reports

Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

doi: 10.1016/j.celrep.2023.112646

Figure Lengend Snippet: (A) Representative immunoblots of 3 independent studies of ITG proteins in Nsi and DAP5-silenced MB-231 cells. Dox induction of shRNAs for 4 days followed by immunoblot analysis of equal protein amounts. (B) Representative immunoblots of 3 independent studies of EMT biomarker protein levels in Nsi and DAP5-silenced 4T1 and MB-231 cells carried out as in (A). (C) Representative immunoblots of 3 independent studies of eIF4E and eIF2α proteins in Nsi and DAP5-silenced MB-231 cells carried out as in (A). (D) Representative immunoblots of 3 independent studies to test requirement for eIF3d cap binding activity for DAP5-dependent mRNAs in MB-231 cells. Cells were small interfering RNA (siRNA) silenced for eIF3d for 1 day and transfected with vectors expressing WT, α5, or α11 cap binding mutants of eIF3d for 2 days, and equal protein amounts analyzed using immunoblot. (E) Matrigel Transwell invasion assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Mean number of invading cells/field with SEM from 3 replicates for each cell line. See also . (F) Cell migration wound healing assays performed with 4T1 and MB-231 cells silenced for 24 h with Nsi or DAP5 Dox-inducible shRNAs. Time 0 represents 100% wound separation of the cell layer. Mean of percentage of migrated cell surface areas (percentage closure) with SEM from 3 replicates for each cell line. See also . (G) Induction of apoptosis by cell detachment (anoikis) determined in 4T1 and MB-231 cells silenced for 24 and 48 h for Nsi, DAP5, or eIF4GI, comparing cells maintained on adherent or ultra- low-adherence plates. Percentage cell apoptosis determined by annexin V-fluorescein isothiocyanate (FITC) staining and flow cytometry, quantified using FlowJo software. Mean with SEM of 3 independent studies per cell line. (H) Representative immunoblot of 3 independent studies of DAP5, eIF4G1, eIF3d, and control β-actin protein levels in Nsi or DAP5-silenced 4T1 and MB-231 cells, silenced for 2 days with Dox-inducible shRNAs. Data are represented as mean ± SEM. n.s., not significant. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 by unpaired parametric two-tailed t test.

Article Snippet: Rabbit polyclonal anti-human/mouse β-actin antibody , Cell Signaling Technology , Cat# 4967/RRID:AB_330288.

Techniques: Western Blot, Biomarker Discovery, Binding Assay, Activity Assay, Small Interfering RNA, Transfection, Expressing, Migration, Staining, Flow Cytometry, Software, Control, Two Tailed Test

Journal: Cell reports

Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

doi: 10.1016/j.celrep.2023.112646

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-human/mouse β-actin antibody , Cell Signaling Technology , Cat# 4967/RRID:AB_330288.

Techniques: Recombinant, Staining, Protease Inhibitor, Virus, Reverse Transcription, SYBR Green Assay, Bicinchoninic Acid Protein Assay, MTT Assay, Software, Imaging

Journal: Cell metabolism

Article Title: Carbon source availability drives nutrient utilization in CD8 + T cells

doi: 10.1016/j.cmet.2022.07.012

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-human/mouse beta-actin , Cell Signaling , #4967.

Techniques: Virus, Luciferase, shRNA, Plasmid Preparation, Recombinant, Staining, Cell Isolation, Selection, Protease Inhibitor, Software

Journal: PLoS ONE

Article Title: Crucial Role of Heme Oxygenase-1 on the Sensitivity of Cholangiocarcinoma Cells to Chemotherapeutic Agents

doi: 10.1371/journal.pone.0034994

Figure Lengend Snippet: ( A ) The basal mRNA expression of HO-1 in CCA cells; KKU-100 and KKU-M214 cells. Total RNA of cells were collected and analysed by real-time RT-PCR. The bars represent relative expression of HO-1 normalized with β -actin. The expression of HO-1 in KKU-100 cells is much higher than that of KKU-M214 cells, p<0.05. ( B ) The basal protein expression of HO-1 in KKU-100 and KKU-M214 cells. Total cell lysates were prepared and subjected to Western blot analysis using β-actin as a loading control. Image samples of HO-1 and β -actin are shown in the top panel of the figure. HO-1 protein in KKU-100 cells is relatively higher than that of KKU-M214 cells. The bars represent mean±SEM, each from three separated experiments. *, p <0.05 vs KKU-100 group.

Article Snippet: The PVDF membrane was incubated overnight at 4°C with primary antibodies of rabbit polyclonal anti-human HO-1 (1∶1000) (ADI-SPA-895: Enzo Life Sciences, Switzerland), rabbit monoclonal anti-human p21 Cip/WAF1 (1∶500) (#2947: Cell Signaling Technology, MA, USA), mouse monoclonal anti-human cytochrome c (1∶1000) (sc-13560) and horseradish peroxidase (HRP)-goat polyclonal anti-human β-actin (1∶2500) (sc-1616 HRP) in TBS.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: PLoS ONE

Article Title: Crucial Role of Heme Oxygenase-1 on the Sensitivity of Cholangiocarcinoma Cells to Chemotherapeutic Agents

doi: 10.1371/journal.pone.0034994

Figure Lengend Snippet: ( A ) KKU-100 cells and ( B ) KKU-M214 cells were cultured for overnight and exposed to Gem (1 µM) for 3, 6, 12 and 24 h. The cultured cells exposed to vehicle only were performed as concurrent control groups. Total cell lysates were collected at the indicated time points and subjected to Western blot analysis using β-actin as a loading control. The relative expression of HO-1 was normalized with β actin. The bars show the HO-1 expression in Gem-treated group and concurrent control group. The bars represent mean±SEM, each from three separated experiments. *, p <0.05 vs concurrent controls.

Article Snippet: The PVDF membrane was incubated overnight at 4°C with primary antibodies of rabbit polyclonal anti-human HO-1 (1∶1000) (ADI-SPA-895: Enzo Life Sciences, Switzerland), rabbit monoclonal anti-human p21 Cip/WAF1 (1∶500) (#2947: Cell Signaling Technology, MA, USA), mouse monoclonal anti-human cytochrome c (1∶1000) (sc-13560) and horseradish peroxidase (HRP)-goat polyclonal anti-human β-actin (1∶2500) (sc-1616 HRP) in TBS.

Techniques: Cell Culture, Western Blot, Expressing

Journal: PLoS ONE

Article Title: Crucial Role of Heme Oxygenase-1 on the Sensitivity of Cholangiocarcinoma Cells to Chemotherapeutic Agents

doi: 10.1371/journal.pone.0034994

Figure Lengend Snippet: ( A ) The time-course of HO-1 induction by SnCl 2 (10 µM) in KKU-100 and KKU-M214 cells. The cells were cultured for overnight and exposed to SnCl 2 for 3, 6 and 24 h before whole cell lysates were collected and HO-1 protein was determined by the Western blotting, using β-actin; as loading control. The HO-1 protein expression in KKU-100 and KKU-M214 cells is relative stable during the first 6 h of exposure to SnCl 2 , whereas high HO-1 expression is evident during 6 to 24 h. The cytotoxicity of Gem (0.1 µM) and Dox (0.5 µM) with or without SnCl 2 (10 µM) was determined in both cell lines after incubation for 24 h. The cell viability ( B ), apoptotic and necrotic cells ( C ) were evaluated by fluorescent dye staining. Data represent mean±SEM, each from three separated experiments. * , p <0.05 vs drug alone.

Article Snippet: The PVDF membrane was incubated overnight at 4°C with primary antibodies of rabbit polyclonal anti-human HO-1 (1∶1000) (ADI-SPA-895: Enzo Life Sciences, Switzerland), rabbit monoclonal anti-human p21 Cip/WAF1 (1∶500) (#2947: Cell Signaling Technology, MA, USA), mouse monoclonal anti-human cytochrome c (1∶1000) (sc-13560) and horseradish peroxidase (HRP)-goat polyclonal anti-human β-actin (1∶2500) (sc-1616 HRP) in TBS.

Techniques: Cell Culture, Western Blot, Expressing, Incubation, Staining

Journal: PLoS ONE

Article Title: Crucial Role of Heme Oxygenase-1 on the Sensitivity of Cholangiocarcinoma Cells to Chemotherapeutic Agents

doi: 10.1371/journal.pone.0034994

Figure Lengend Snippet: The mRNA (A) and protein (B) levels of HO-1expression in KKU-100 cells are shown. KKU-100 cells were transfected with siRNA against HO-1 for 24 h and total RNA was prepared and analyzed by reverse transcription-PCR. In similar experiments, cell lysates were collected and HO-1 was determined by the Western blotting in KKU-100 cells, using β-actin, as loading control. ( C ) The cytotoxicity and IC50 of Gem in knocked down KKU-100 cells was determined. After transfection for 24 h, KKU-100 cells were treated with varied concentrations of Gem (0.001, 0.01, 0.05 and 0.1 µM) for another 24 h. The cell viability was evaluated by fluorescent dye staining. Data represent mean±SEM, each from three separated experiments. * , p <0.05 vs non target knocked down cells.

Article Snippet: The PVDF membrane was incubated overnight at 4°C with primary antibodies of rabbit polyclonal anti-human HO-1 (1∶1000) (ADI-SPA-895: Enzo Life Sciences, Switzerland), rabbit monoclonal anti-human p21 Cip/WAF1 (1∶500) (#2947: Cell Signaling Technology, MA, USA), mouse monoclonal anti-human cytochrome c (1∶1000) (sc-13560) and horseradish peroxidase (HRP)-goat polyclonal anti-human β-actin (1∶2500) (sc-1616 HRP) in TBS.

Techniques: Transfection, Western Blot, Staining

Journal: PLoS ONE

Article Title: Crucial Role of Heme Oxygenase-1 on the Sensitivity of Cholangiocarcinoma Cells to Chemotherapeutic Agents

doi: 10.1371/journal.pone.0034994

Figure Lengend Snippet: The Western blots of p21 and cytochrome c protein expression in KKU-100 cells after treatment with Gem (0.01 µM)+/−ZnPP (0.1 µM) for 24 h. The levels of p21 and cytochrome c were normalized with β-actin as a loading control. The bars are mean±SEM, each from three independent experiments. * p <0.05 vs untreated controls.

Article Snippet: The PVDF membrane was incubated overnight at 4°C with primary antibodies of rabbit polyclonal anti-human HO-1 (1∶1000) (ADI-SPA-895: Enzo Life Sciences, Switzerland), rabbit monoclonal anti-human p21 Cip/WAF1 (1∶500) (#2947: Cell Signaling Technology, MA, USA), mouse monoclonal anti-human cytochrome c (1∶1000) (sc-13560) and horseradish peroxidase (HRP)-goat polyclonal anti-human β-actin (1∶2500) (sc-1616 HRP) in TBS.

Techniques: Western Blot, Expressing