primary antibodies against total erk terk 9102s  (Cell Signaling Technology Inc)

Journal: Biological Research

Article Title: CCAT2 is an oncogenic long non-coding RNA in pancreatic ductal adenocarcinoma

doi: 10.1186/s40659-017-0149-0

Figure Lengend Snippet: KRAS regulates CCAT2 expression via MEK/ERK pathway. a The expression of KRAS was compared between Scramble- and si-KRAS-treated PANC-1 cells 48 h after transfection. b CCAT2 expression in PANC-1 cells transfected with si-KRAS or Scramble. *P < 0.05. c Immunoblotting for p-ERK, total ERK, p-AKT and total AKT in si-KRAS-treated and Scramble-treated PANC-1 cells 48 h after transfection. d PANC-1 cells were co-transfected with CCAT2 promoter-reporter plasmid in combination with si-KRAS or Scramble. Luciferase activity were measured 48 h after transfection. *P < 0.05. For a and c , GAPDH was used as loading control. Image shown was representative of three independent biological repeats. For b and d , data shown were mean ± SEM of three independent biological repeats. e Activation of ERK or AKT was evaluated in PANC-1 cells treated with MEK/ERK inhibitor PD98059 or PI3K/AKT inhibitor LY294002 for 24 h. f CCAT2 expression in PANC-1 cells treated with PD98059 or LY294002 for 24 h. *P < 0.05. For e and f , DMSO-treated group was used as control. Image shown was representative of three independent biological repeats

Article Snippet: Primary rabbit antibodies against total ERK (No. 4695), p-ERK (No. 4370), total AKT (No. 4685), p-AKT (No. 4060), KRAS (No. 3339) (Cell Signaling Technology Inc., Berkeley, CA, USA) and GAPDH (No. TA-08) (loading control) (Zhongshanjinqiao Biotech, China) were incubated at 4 °C overnight at a dilution of 1:1000, and after washed with PBST for three times, the secondary horseradish-peroxidase-labeled antibody was incubated at room temperature for 2 h at a dilution of 1:5000.

Techniques: Expressing, Transfection, Western Blot, Plasmid Preparation, Luciferase, Activity Assay, Control, Activation Assay

Journal: PLoS ONE

Article Title: Generation and characterization of Kctd15 mutations in zebrafish

doi: 10.1371/journal.pone.0189162

Figure Lengend Snippet: Total-ERK (tERK) antibody staining of (A) wild-type and (B) mutant 6 dpf larvae. C) Pixels whose intensity values are statistically significantly different between wildtype and mutant brains (p < 0.05, N = 14 per group, see for statistical comparison procedure). Sectioning and H+E staining of a large region of adult (5 month old) wild-type (D) and mutant (E) brains revealed that this structure (green square in D) is absent throughout development. At least fifteen larval brains and five adult brains from each genotype were analyzed to confirm these results.

Article Snippet: Larvae at 6 dpf were fixed in 4% PFA and labeled using an anti-tERK primary antibody (1:500, Cell Signaling, 4696) and Alexa488 secondary antibody.

Techniques: Staining, Mutagenesis, Comparison