The process of immunofluorescence analysis was performed as described by our previous studies26 (link). Briefly, brain slices were permeabilized with 0.3% Triton-X 100 and 5% BSA in PBS and incubated with primary antibodies at 4 °C overnight. After washing with PBS three times, samples were incubated with Alexa Fluor-555 secondary antibodies (goat-anti-mouse, A32727; goat-anti-rabbit, A32732; 1:1000, Thermo Fisher) for 1 h at room temperature. Samples were mounted with Vectashield mounting medium (Vector Labs) and images were taken by Leica TCS SP8 confocal microscope. The following primary antibodies were used: rabbit anti-NeuN (1:500, Abcam, ab177487), mouse anti-GABA (1:1000, Invitrogen, PA5-32241), and rabbit anti-neurogranin (1:1000, R&D, MAB7947). Unbiased stereology TissueFAX Plus ST (Tissue Gnostics, Vienna, Austria)73 (link) was applied to count EGFP-positive and NeuN-positive cell number in brain slices.
Ab177487
Manufactured by Thermo Fisher Scientific
Ab177487 is a laboratory instrument designed for the detection and analysis of specific biological molecules. The core function of this product is to provide accurate and reliable measurements of target analytes in various sample types. Detailed specifications and intended use are not available for this particular product.
Automatically generated - may contain errors
Lab products found in correlation
A32732, by Thermo Fisher Scientific (1 mentions)
A32727, by Thermo Fisher Scientific (1 mentions)
Tcs sp8 confocal microscope, by Leica (1 mentions)
Vectashield mounting medium, by Vector Laboratories (1 mentions)
Anti v5, by Thermo Fisher Scientific (1 mentions)
R960 25, by Thermo Fisher Scientific (1 mentions)
Ab177487, by Abcam (1 mentions)
Anti arl13b, by Proteintech (1 mentions)
Anti pericentrin, by Abcam (1 mentions)
Anti pericentrin, by BD (1 mentions)
Anti arl13b, by Abcam (1 mentions)
Anti neun, by Abcam (1 mentions)
2 protocols using ab177487
1
Immunofluorescence Analysis of Brain Slices
2
Immunolabeling Neuronal Markers and Receptors
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We used the following antibodies: Anti-ARL13B (IF:1/500, western blot (WB): 1/1000; NeuroMab, 11000053); Anti-HA (clone 3F10, IF:1/1000, 11867423001, Sigma); Anti-NeuN (IF: 1:300, ab177487, Abcam); Anti-V5 (IF: 1:500, R960-25, Invitrogen); Anti-DR6 (TNFRSF21) (clone 6B6, IF: 1:200, MABC1594-25UL, Millipore); Anti-ARL13B (Immunofluorescence (IF): 1:500, Abcam, ab136648); Anti-ARL13B (IF: 1:500, Proteintech, 17711-11AP); Anti-Pericentrin (IF: 1:500, BD Biosciences, 611814); Anti-Pericentrin (IF: 1:500, Abcam, ab4448); Anti-NeuN (IF: 1:500, Abcam, ab104224); Anti-NeuN (IF: 1:500, Abcam, ab177487); Anti-GFP (IF: 1:1000, Invitrogen, A11122); Anti-V5 Tag (IF: 1:500, Invitrogen, R960-25); Anti-Adenylate Cyclase 3 (IF: 1:500, Invitrogen, PA5-35382); Anti-Ephrin-B1 (IF: 1:50, Bio-Techne, AF473); Anti-Ephrin-B2 (IF: 1:50, Bio-Techne, AF496); Anti-EphB2 (IF: 1:50, Bio-Techne, AF467); Anti-EphA4 (IF: 1:50, Proteintech, 21875-1-AP); Anti-CSNK1G1 (IF: 1:50, OriGene, TA806333S); Anti-ROBO2 (IF: 1:100, GeneTex, GTX134119); Anti-UNC5C (IF: 1:100, LSBio, LS-B8305-50); Anti-SEMA6B (IF: 1:100, biorbyt, orb422749); Anti-SLC6A1 (IF: 1:500, Invitrogen, PA5-85766); Anti-GABRG2 (IF: 1:100, Proteintech, 14104-1-AP); Anti-GABBR1 (IF: 1:500, Invitrogen, PA5-27725); Anti-GABRA1 (IF: 1:100, Proteintech, 12410-1-AP), Anti-DR6 (Tnfrsf21) (IF: 1:100, Sigma, MABC1594).
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