The antigen peptides for all LRIG1 antibodies used are indicated in Supplementary Fig. 1B . For immunohistochemistry: Anti-LRIG1 antibody (Atlas Antibodies AB, Bromma, Sweden, catalog no. HPA011846), 4.0 mg/ml. The 1A8 anti-LRIG1 antibody is described in the Supplementary Materials. For western blots: EGFR #2232, 1:1000; Phospho-p44/42 MAPK #4370, 1:1000; Phospho-AKT #4060, 1:1000 (Cell Signaling Technology Inc, Danvers, USA); Anti-Actin #ACTN05/C4, 1:3000 (Abcam, Cambridge, UK). Anti-LRIG1 Vina [49 (link)], 1:1000 (Agrisera AB, Vännäs, Sweden). Secondary antibodies: IRDye® 680RD Donkey anti-Mouse #926-68072, 1:15000; IRDye® 800CW Goat anti-Rabbit #926-32211, 1:15000 (LI-COR, Lincoln, NE, USA).
Anti actin actn05 c4
Manufactured by Abcam
Sourced in United States, United Kingdom
Anti-Actin #ACTN05/C4 is a monoclonal antibody that specifically recognizes the actin protein. Actin is a ubiquitous and highly conserved cytoskeletal protein found in all eukaryotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Phospho akt 4060, by Cell Signaling Technology (1 mentions)
Irdye 800cw goat anti rabbit 926 32 211, by LI COR (1 mentions)
Image lab v6, by Bio-Rad (1 mentions)
Anti p44 42 mapk erk1 2, by Cell Signaling Technology (1 mentions)
Ripa buffer, by Thermo Fisher Scientific (1 mentions)
Bca assay, by Thermo Fisher Scientific (1 mentions)
Precast gel, by Bio-Rad (1 mentions)
Anti phosp ho 4e bp1 ser65, by Cell Signaling Technology (1 mentions)
Complete mini edta free protease inhibitor cocktail, by Roche (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
Supersignal west femto maximum sensitivity substrate, by Bio-Rad (1 mentions)
Anti phospho p44 42 mapk erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
2 protocols using anti actin actn05 c4
1
Antibody Characterization for LRIG1 Analysis
2
Protein Expression Analysis in Drug Treatments
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Protein extracts were obtained after six days of incubation with each drug alone or in combination. For this purpose, RIPA Buffer (Thermo Fisher Scientific) containing cOmplete™, Mini, EDTA-free Protease Inhibitor Cocktail (Roche Applied Science) and Phosphatase Inhibitor Cocktail (Active Motive) was used. Protein concentration was determined with a BCA assay (Thermo Fisher Scientific). Then, proteins were separated by SDS-PAGE on precast gels (Bio-Rad Laboratories, Hercules, CA, USA), transferred to PVDF membranes and incubated overnight at 4ᵒC with the correspondent primary antibody. Next day, membranes were incubated with HRP-conjugated secondary antibodies for 1 h at room temperature. Peroxidase activity was detected using the SuperSignal™ West Femto Maximum Sensitivity Substrate (Bio-Rad). Images were captured with a ChemiDoc™ MP Imaging System and analyzed with Image Lab™ v6 software (Bio-Rad). ImageJ was used for densitometry analysis. The following antibodies were used: anti-actin [ACTN05 (C4)] (Cat# ab3280) from Abcam (Cambridge, UK); anti-4E-BP1 (53H11) (Cat#9644), anti-Phospho-4E-BP1 (Ser65) (Cat#9451), anti-p44/42 MAPK (ERK1/2) (Cat#9102) and anti-Phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (Cat#9101) from Cell Signalling Technology (Danvers, MA, USA).
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