Monoclonal antibodies were against GPP130 (Linstedt et al., 1997 (link)), myc (Evan et al., 1985 (link); Jesch et al., 2001 (link)), HA (H3663, Clone HA-7; Sigma), giantin (Linstedt and Hauri, 1993 (link)), GFP (SAB2702197, clone GT859; Sigma), sortilin (Cat# 612100, clone 48/Neurotensin; BD Biosciences), and tubulin (T6557, clone GTU-88; Sigma). Polyclonal antibodies were against GPP130 (Puri et al., 2002 (link)), TMEM165 (NBP1-90651; Novus Biologicals), GAPDH (14C10; Cell Signaling Technology), and sortilin (a kind gift from Claus M. Petersen, Aarhus University [ Petersen et al., 1997 (link)]). Secondary antibodies were Alexa 488 anti-mouse (Cat#A28175; Thermo Fisher Scientific), Alexa 488 anti-rabbit (Cat#A27034; Thermo Fisher Scientific), Alexa 555 anti-rabbit (Cat#A27039; Thermo Fisher Scientific), Alexa 555 anti-mouse (Cat#A28180; Thermo Fisher Scientific), and horseradish peroxidase–conjugated goat anti-mouse (Cat#170-6516; Sigma) and goat anti-rabbit antibodies (Cat#170-6515; Sigma). AP12998 was from Clontech (called D/D solubilizer, Cat#635054). Cycloheximide (Cat#C7698) and monensin (Cat#M5273) were from Sigma. ECL blotting substrate (Cat#32209) and MnCl2 (Cat#M87-500) were from Thermo Fisher Scientific.
Sortilin
Manufactured by Cell Signaling Technology
Sortilin is a type I transmembrane protein that functions as a sorting receptor, involved in the intracellular trafficking and localization of various proteins and ligands. It plays a role in the endocytic and lysosomal pathways.
Automatically generated - may contain errors
Lab products found in correlation
Sortilin, by BD (1 mentions)
Monensin, by Merck Group (1 mentions)
Ecl blotting substrate, by Thermo Fisher Scientific (1 mentions)
Mncl2, by Thermo Fisher Scientific (1 mentions)
Anti mouse alexa 488, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit antibodies, by Merck Group (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Glut4, by Cell Signaling Technology (1 mentions)
P akt ser473, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
2 protocols using sortilin
1
Antibody Characterization and Experimental Conditions
2
Western Blot Analysis of Insulin Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cell lysates (50 µg) were harvested using lysis buffer (Cell Signaling 9803S) + 10% protease/phosphatase inhibitor (Pierce A32957, A32953) then sonicated briefly. Samples were separated on a 7% SDS-PAGE gel. Proteins were electrophoretically transferred to nitrocellulose membranes and blocked with 5% nonfat dried milk in Tris-Buffered Saline with 0.05% Tween 20 (TBST). Membranes were probed with Sortilin (ab16640), Glut4 (Cell Signaling #2213), pAkt (Ser 473, Cell Signaling #4058), Akt2 (Cell Signaling #2962), and β-Actin (Sigma A3854). Secondary HRP antibodies were purchased from Biorad for rabbit (5196-2504) and mouse (0300-0108P). Incubation with chemiluminesence (Pierce 32109) was used for detection and images were digitally captured using ProteinSimple FluorChem MTM and densitometric analysis was performed using AlphaView Software.
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