hES-MN cultures (day 35) were fixed in 4 % paraformaldehyde-PBS for 10 min and permeabilized with 0.1 % Triton X-100. After being blocked with 10 % serum, the cells were then incubated overnight with the following primary antibodies: Tau5 (Thermo Scientific), LIM3 (Millipore), ChAT (Millipore), NeUN (Millipore), Hb9 [Developmental Studies Hybridoma Bank (DSHB)], Isl1 (DSHB), and β-III tubulin (Covance) (as shown in Fig. 1b ), as well as antibodies for SNAP-25 (BD Biosciences), SV2A (Millipore), Syntaxin (Sigma), VAMP-2 (R&D Systems), and MAP2 (Millipore) (as shown in Fig. 2b ) in PBS with 10 % serum according to the manufacturers’ suggested working concentrations. On the following day, appropriate secondary antibodies, conjugated with Alexa488 and Alexa594, were incubated with the cells for 2 h at room temperature. Image acquisition was performed using Zeiss or Opera (PerkinElmer) confocal microscopes.
Snap 25
Manufactured by BD
SNAP-25 is a protein that plays a key role in the regulation of synaptic vesicle exocytosis, a process essential for neurotransmitter release. It is a component of the SNARE complex, which facilitates the fusion of synaptic vesicles with the presynaptic membrane. SNAP-25 is a widely used marker for detecting and measuring synaptic function in various experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Syntaxin, by Merck Group (1 mentions)
Neuronal nuclei (neun), by Merck Group (1 mentions)
Vamp 2, by R&D Systems (1 mentions)
βiii tubulin, by Fortrea (1 mentions)
Transferrin, by Thermo Fisher Scientific (1 mentions)
Psd95, by Cell Signaling Technology (1 mentions)
Glun2b, by Alomone (1 mentions)
Darpp32, by Abcam (1 mentions)
Biii tubulin, by Merck Group (1 mentions)
Calnexin, by Stressgen (1 mentions)
Vglut1, by Synaptic Systems (1 mentions)
Vglut2, by Synaptic Systems (1 mentions)
2 protocols using snap 25
1
Immunocytochemical Characterization of hES-MN Cultures
2
Comprehensive Neurological Protein Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Actin (Sigma, A4700) 1:400, Alpha Actinin 2 (Abcam, ab68167) 1:2000, AMPA Receptor 1 GluR1a (Alomone, AGC-004) 1:500, BIII Tubulin (Sigma, T8660) 1:2000, Calnexin (StressGen, SPA-860) 1:2000, DARPP32 (Abcam, 40801) 1:10,000, GABA(A) a1 Receptor (Alomone, AGA-001) 1:500, Glucose Transporter GLUT3 (Abcam, ab41525) 1:500, Na+/K+ ATPase (Affinity Bioreagents, MA3-915) 1:5,000, NMDA Receptor 2B GluN2B (Alomone, AGC-003) 1:500, PDE10a (Abcam, 177933) 1:2000, PSD95 (Cell Signaling, 2507S) 1:1000, SCN4B (Abcam, ab80539) 1:200, SNAP25 (BD Transduction Labs, 610366) 1:10,000, Transferrin (Thermo Fisher, 13-6800) 1:1000, VGlut1 (Synaptic Systems, 135302) 1:10,000, VGlut2 (Synaptic Systems, 135402) 1:10,000, XK (Aviva Systems Bio, ARP33809_P050) 1:1000, Huntingtin [Ab1, aa1-17, (DiFiglia et al., 1995 (link))] 1:2000, poly-Q (1C2, EMD Millipore MAB1574) 1:1000. Horseradish peroxidase secondary antibodies (Jackson Immunoresearch) were diluted 1:5000.
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