The unroofed cells were incubated in 3% bovine serum albumin/PBS (BSA, m/v, fresh, Fisher Bioreagents #BP9703) for 1.5 h, followed by primary antibody (1:100 in 3%BSA/PBS) incubation for 1 h. Next, cells were washed thoroughly with PBS and the secondary antibody tagged with fluorescence dye (1:500) or GFP-nanobody (1:500) was applied for 1 h. Afterwards, cells were washed 4 times in PBS and stored in fresh PBS at 4 C until the samples were imaged. The following antibodies were used: anti-Caveolin1-Rabbit (abcam #ab2910), anti-Caveolin1-mouse (Santa Cruz #sc-53564), anti-Cavin1-Rabbit (abcam #76919), anti-Cavin2-Rabbit (abcam #ab76867), anti-Cavin3-Rabbit (abcam #abcam2912), anti-EHD2-goat (abcam #ab23935), anti-Pacsin2-Rabbit (Proteintech #10518-2-AP), anti-EHBP1-Rabbit (Proteintech #17637-1-AP), anti-mouse-Clathrin heavy chain (Thermo-Fisher #MA1-065, 1:2000), anti-mouse-Dynamin2 (Santa Cruz, C-18; #sc-6400), anti-rabbit-Atto647N (Rockland #611-156-122), anti-goat-Atto647N (Rockland, #610-156-121), anti-goat-Atto647N (Rockland #605-456-013 S), anti-rabbit-Alexa568 (Invitrogen #A11036), Fab2-anti-rabbit-Alexa594 (ThermoFisher #A-11072), Fab2-anti-mouse-Alexa488 (ThermoFisher #A-11017), Fab2-anti-mouse-Alexa568 (ThermoFisher #A-11019), GFP-nanobody-Atto647N (Chromotek #gba647n-100), Phalloidin-Alexa488 (ThermoFisher #A12379).
Ab2910
Manufactured by Santa Cruz Biotechnology
Ab2910 is a laboratory instrument used for the purification and isolation of proteins and other biomolecules. It utilizes affinity chromatography techniques to separate and purify target molecules from complex mixtures. The core function of Ab2910 is to provide a reliable and efficient method for the preparation of high-purity samples for further analysis and research purposes.
Automatically generated - may contain errors
2 protocols using ab2910
1
Immunofluorescence Staining of Caveolins
2
Insulin-Induced Cav1-Insr Interaction
Check if the same lab product or an alternative is used in the 5 most similar protocols
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MCECs were seeded at low density onto coverslips and serum‐starved overnight. Then, they were incubated with 100 nM insulin for the indicated period, washed, and fixed with 4% PFA before they were blocked for 1 h in 5% BSA in TBST. PLA was performed with primary antibodies against Cav1 (abcam ab2910) and Insr beta (Santa Cruz sc‐31367). PLA was performed according to the manufacturer's guidelines (Duolink Orange Detection System, Olink Biosciences). PLA spots were counted with a fluorescence widefield microscope.
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