Immunoblotting Analysis of ER Stress Markers

Primary antibodies against PDI (rabbit monoclonal; 1:1000), Ero1-Lα (rabbit monoclonal; 1:1000), IRE1α (rabbit monoclonal; 1:1000), Calnexin (rabbit monoclonal; 1:600), eIF2α (rabbit polyclonal; 1:800), P-eIF2α (rabbit monoclonal; 1:600), and CHOP (mouse monoclonal; 1:800) were purchased from Cell Signaling Technology, Frankfurt am Main, Germany. Antibodies NADPH 4 oxidase (rabbit polyclonal; 1:800), and Total OXPHOS cocktail antibody (rodent monoclonal; 1:800) were purchased from Abcam, Cambridge, UK. Hsp70 (mouse monoclonal; 1:800), and HO-1 (mouse monoclonal; 1:800) were purchased from Enzo life sciences, Lörrach, Germany. β-actin (rabbit polyclonal; 1:1200) from Novusbio, Centennial, CO, USA, and Vinculin (mouse monoclonal, 1:500) from AbD Serotec, Puchheim, Germany. Where applicable secondary antibodies used during immunoblotting were Donkey anti rabbit (1:8000; Novex), Horse anti mouse (1:2000; Cell Signaling Technology), Donkey anti mouse (1:5000; Bethyl), Goat anti rabbit (1:2000; Cell Signaling Technology), and Donkey anti rabbit (1:4000; Novex).

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Bola S., Subramanian P., Calzia D., Dahl A., Panfoli I., Funk R.H, & Roehlecke C. (2023). Analysis of Electric Field Stimulation in Blue Light Stressed 661W Cells. International Journal of Molecular Sciences, 24(4), 3433.

Publication 2023

Ero1-Lα IRE1α Calnexin eIF2α P-eIF2α Total OXPHOS cocktail antibody Hsp70 β-actin Donkey anti rabbit Horse anti mouse Goat anti rabbit

Actin Antibodies Antibody Calnexin Chop Donkey Goat Horse Hsp70 Ire1α Mouse Nadph oxidase Rabbit Rodent Vinculin

Corresponding Organization : TU Dresden

Other organizations : University of Genoa

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Variable analysis

independent variables

Primary antibodies against PDI (rabbit monoclonal; 1:1000)
Primary antibodies against Ero1-Lα (rabbit monoclonal; 1:1000)
Primary antibodies against IRE1α (rabbit monoclonal; 1:1000)
Primary antibodies against Calnexin (rabbit monoclonal; 1:600)
Primary antibodies against eIF2α (rabbit polyclonal; 1:800)
Primary antibodies against P-eIF2α (rabbit monoclonal; 1:600)
Primary antibodies against CHOP (mouse monoclonal; 1:800)
Primary antibodies against NADPH 4 oxidase (rabbit polyclonal; 1:800)
Primary antibodies against Total OXPHOS cocktail (rodent monoclonal; 1:800)
Primary antibodies against Hsp70 (mouse monoclonal; 1:800)
Primary antibodies against HO-1 (mouse monoclonal; 1:800)
Primary antibodies against β-actin (rabbit polyclonal; 1:1200)
Primary antibodies against Vinculin (mouse monoclonal, 1:500)
Secondary antibodies (various types and concentrations)

dependent variables

Protein expression levels of PDI, Ero1-Lα, IRE1α, Calnexin, eIF2α, P-eIF2α, CHOP, NADPH 4 oxidase, Total OXPHOS, Hsp70, HO-1, β-actin, and Vinculin

control variables

Concentration of primary antibodies
Concentration of secondary antibodies
Experimental conditions (e.g., cell type, treatment, etc.)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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