The following antibodies were used: VAMP2 rabbit antibody (#104202; Synaptic System), SV2 mouse antibody (Developmental Studies Hybridoma Bank), Flag M2 mouse antibody (#1804; Sigma-Aldrich), tubulin mouse antibody (#t9026; Sigma-Aldrich), mCherry rabbit antibody (#5993; BioVision), Myc mouse antibody (9E10; Santa Cruz), Hrs rabbit antibody (D7T5N; Cell Signaling), Hrs mouse antibody (ab56468; Abcam), GFP mouse antibody (#11814460001; Roche), KIF13A rabbit antibody (PA5-30874; Thermo Fisher Scientific), KIF13B mouse antibody (6E11; Santa Cruz), neurofilament and β-actin mouse antibodies (2H3 and 8-7A5; Developmental Studies Hybridoma Bank). Pharmacological agents were used in the following concentrations and time courses: cycloheximide (Calbiochem, 0.2 μg/μl, 24 h), bicuculline (Sigma-Aldrich, 40 μM, 5 min-2h or 20 h), 4-aminopyridine (Tocris Bioscience, 50 μM, 5 min-2h or 20 h), BDNF (R&D Systems, 50 ng/ml, 1 h), TTX (Tocris Bioscience, 0.5 μM, 2 h), Rapamycin analogue linker drug (AP21967; Clontech, 100 nM, 3 h), SAR405 (Millipore Sigma-Aldrich, 1 μM, 24 h), and Janelia Fluor 549 SNAPtag ligand (Janelia Research Campus, 200 nM, 30 min). Unless otherwise indicated, all other chemicals were purchased from Sigma-Aldrich.
Flag m2 mouse antibody
Manufactured by Merck Group
The Flag M2 mouse antibody is a laboratory reagent used for the detection and purification of proteins that have been engineered to contain a specific amino acid sequence known as the Flag epitope. The antibody binds to this epitope, allowing for the identification and isolation of the target protein from complex biological samples.
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Lab products found in correlation
2 protocols using flag m2 mouse antibody
1
Antibodies and Pharmacological Agents for Neuronal Imaging
2
Immunofluorescence Staining of FLAG-tagged Proteins
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Cells were seeded in 3.5-cm glass bottom dishes. When the cell density reached about 80% confluency, the cells were washed with PBS, and fixed with 4% formaldehyde in PBS for 30 min. Then the cells were washed with PBS for 5 min, and treated with 0.2% Triton X-100 in PBS for 2 min to enhance antibody penetration. The cells were blocked at room temperature for 15 min with 10% goat serum in TN buffer (0.1 M Tris-HCl pH 7.5, 0.15 M NaCl), and then incubated with FLAG M2 mouse antibody (Sigma) in TNT buffer (TN buffer containing 0.05% Tween 20) for 1 hour at room temperature. The cells were washed 3 times with PBS and treated with Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Invitrogen) in TNT buffer at room temperature for 1 hour. Then fluorescence was observed using a fluorescence microscope (LX70, Olympus).
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