Concentrations of fusion proteins were measured on a ND-1000 Nanodrop spectrophotometer (Thermo Scientific) by UV-vis absorption spectroscopy. Concentration of exendin and conjugates for in vitro assays and in vivo studies was assessed using a Bicinchoninic Acid (BCA, Pierce) assay following manufacturer’s instructions. SDS-PAGE analysis of sortase A was performed using precast 4–20% Tris-HCl gels (Bio-Rad). SDS-PAGE analyses of all exendin derivatives were performed using precast Tris/Tricine gels (Bio-Rad). Quantification of sortase reaction conversion was done by gel densitometry analysis using a built-in function in Image Lab (v. 4.0.1, Bio-Rad).
Precast tris tricine gels
Manufactured by Bio-Rad
Precast Tris/Tricine gels are electrophoresis gels that are pre-cast and ready-to-use. They are designed for the separation and analysis of proteins, particularly smaller proteins and peptides, using the Tris/Tricine buffer system.
Automatically generated - may contain errors
Lab products found in correlation
Image lab v4, by Bio-Rad (2 mentions)
Nanodrop nd 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Precast 4 20 tris hcl gels, by Bio-Rad (2 mentions)
Image studio v2, by LI COR (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Superdex 200 10 300 gl column, by GE Healthcare (1 mentions)
4 protocols using precast tris tricine gels
1
Quantitative Analysis of Exendin Derivatives
2
Histone Acetylation and SWI/SNF Complex Analysis
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Cytosolic and nuclear protein fractions were isolated using a commercially available kit (Cell Biolabs, San Diego, CA), and histones were extracted from the remaining nuclear pellet by 1 min sonication in a sonicating water bath. Histones were separated with pre-cast tris-tricine gels (Bio-Rad) then transferred to a PVDF membrane for blotting. All primary acetyl histone antibodies were used at a 1:500 in 4 % BSA-TBS (0.01 % Tween-20). For analysis of SWI/SNF proteins 30 μg nuclear protein were loaded into pre-cast 4–20 % tris-glycine gels before being transferred to a PVDF membrane for blotting. All SWI/SNF antibodies were from a SWI/SNF complex antibody sampler kit (Cell Signal Technology) and were used at 1:1000 in 5 % NFDM-TBS (0.01 % Tween-20). All experiments and blots were performed at least in triplicate and relative expression was quantified using an Odyssey Infrared Imaging system and Image Studio v. 2.0 software (LI-COR, Lincoln, NE) using total Histone H3 protein as the invariant control.
3
Quantitative Analysis of Exendin Derivatives
4
CSF Amyloid-beta Fractionation and Analysis
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Five hundred μL of CSF from a normal individual was depleted of endogenous IgGs and loaded onto a Superdex 200 10/300 GL column (GE Healthcare) and eluted with 50 mM ammonium acetate, pH 8.5 at a flow rate of ~0.3 mL/min. Fractions of 250 μL were collected, and 125 μL aliquots of each selected fraction were analyzed by WB. Specifically, proteins were separated in 10–20% Precast Tris-Tricine gels (Bio-Rad), electrophoretically transferred onto nitrocellulose membranes, and probed using biotinylated 6E10 or anti-Aβx-40/42.
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