These materials using in this study are mentioned in prior study, such as 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), DMSO 5-FU, RIPA lysis buffer, crystal violet, 4% paraformaldehyde solution, Dulbecco’s modified eagle medium (DMEM), penicillin-streptomycin, Phosphate buffered saline (PBS), Fetal Bovine Serum (FBS), Pierce BCA protein assay kit, and Super Signal™ West Pico Chemiluminescent Substrate kit, TRIzol lysis buffer, and The Prime Script RT reagent Kit and TB Green™ Premix Ex Taq™ II (Tli RNaseH Plus) [10 ,14 ]. Hoechst 33258 reagent was from Beyotime Biotechnology (Jiangsu, China). The primary antibodies against Bcl-2, Bax, cleaved-caspase 3, cleaved caspase 9, GAPDH and β-actin were purchased from Cell Signaling Technology (Beverly, MA, USA). P-R-Ras, P-ERK1/2, P-PI3K, VEGFA, P-AKT, HIF-1A, and P-ERK1/2 antibodies were purchased from Affinity Biosciences (Cincinnati, OH, USA).
P erk1 2
Manufactured by Affinity Biosciences
Sourced in United States
P-ERK1/2 is a lab equipment product that measures the phosphorylation levels of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) proteins. ERK1/2 are key components of the mitogen-activated protein kinase (MAPK) signaling pathway, which plays a crucial role in the regulation of various cellular processes, such as cell growth, differentiation, and survival. The P-ERK1/2 product provides a quantitative assessment of the activation state of the ERK1/2 proteins, which can be used as an indicator of the activity of the MAPK pathway in different biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Erk1 2, by Affinity Biosciences (2 mentions)
P akt, by Affinity Biosciences (1 mentions)
Hoechst 33258 reagent, by Beyotime (1 mentions)
Hif 1a, by Affinity Biosciences (1 mentions)
Vegfa, by Affinity Biosciences (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Cleaved caspase 9, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
P pi3k, by Affinity Biosciences (1 mentions)
Dlk 1, by Abcam (1 mentions)
Horseradish peroxidase hrp labeled secondary antibody, by Proteintech (1 mentions)
Gapdh, by Merck Group (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Cebp α, by Affinity Biosciences (1 mentions)
P smad1 5 8, by Affinity Biosciences (1 mentions)
P acc, by Sangon (1 mentions)
P smad2 3, by Affinity Biosciences (1 mentions)
4 protocols using p erk1 2
1
Comprehensive Cell Signaling Protocol
2
Dlk1 Overexpression Protein Analysis
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The total protein content was extracted from cells of each cell line by using RIPA lysis buffer and then determined by using the BCA method. The whole protein of each sample was separated using 10% SDS-PAGE and transferred onto PVDF membranes for blotting. The antibodies used in this study were Gapdh (#60004-1-Ig Proteintech, Wuhan, China), Dlk1 (#ab210471, Abcam, Cambridge, UK), Notch1 (#4ab094535, 4A BIOTECH, Beijing, China), Rhoc (#4ab090482, 4A BIOTECH, Beijing, China), Erk1/2 (#BF8004, Affinity Biosciences, Changzhou, China), and p-Erk1/2 (#AF1015, Affinity Biosciences, Changzhou, China). The full length of Dlk1 cDNA was cloned to pcDNA3.1 to overexpress Dlk1.
3
Quantification of Cardiac Fibroblast Proteins
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The total proteins were isolated from cardiac fibroblasts that were western blotted using the monoclonal antibodies against EGFR (1 : 1, 000, Abcam, Cambridge, MA, USA), ERK1/2 (1 : 1, 000, Affinity, USA), and p-ERK1/2 (1 : 1, 000, Affinity). The loading control was served by GAPDH (1 : 1, 000, Sigma-Aldrich, USA). Cardiac fibroblasts were cultured at 25°C for 1 h with a horseradish peroxidase (HRP) labeled secondary antibody (1 : 1,000, Proteintech, Chicago, USA). Band density quantifications were investigated using an Odyssey Infrared Imaging System (LI-COR Biosciences, Lincoln, NE, USA).
4
Comprehensive Signaling Pathway Analysis
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Affinity Biosciences: p-SMAD1/5/8 at Ser463 + Ser465 (AF8313; 1:1000), SMAD1/5/8 (AF0614; 1:1000); p-SMAD2/3 at Thr8 (AF3367; 1:1000); SMAD2/3 (AF6367; 1:1000); C/EBPα (AF6333; 1:1000); p-ERK1/2 at Thr202/Tyr204 (AF1015; 1:1000); ERK1/2 (AF0155; 1:1000); p-p65 at Ser536 (AF2006; 1:1000); p65 (AF5006; 1:1000); p-IKKα/β at Ser180/Ser181 (AF3013; 1:1000); IKKα/β (AF6014; 1:1000). Sangon Biotech: p-ACC at Ser79 (D155180; 1:1000); ACC (D155300; 1:1000). Abcam: BMP8B (ab181253; 1:1000). CWBIO: Goat Anti-Rabbit IgG, HRP Conjugated (CW0103S; 1:4000). Bioss: PPARγ (bs-4590R; 1:1000); p-p38 MAPK at Thr180 + Tyr182 (bs-2210R; 1:1000); p38 MAPK (bs-0637R; 1:1000); β-actin (bs-0061R; 1:2000); p-JNK at Thr183 + Tyr185 (bs-1640R; 1:1000); JNK (bs-2592R; 1:1000).
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