Immunocytochemistry was performed as follows: fixation with 4% paraformaldehyde (PFA) for 20 min in RT; washing with phosphate buffered saline (PBS) twice; permeabilization and blocking in PBS containing 0.1% Triton X-100 and 3% FBS in PBS for 30 min at RT; incubation with primary antibody diluted 1:500 in blocking solution for 2 h in RT; washing with PBS three times; incubation with Hoechst 33258 (Nacalai Tesque) and secondary antibody diluted 1:500 in PBS for 1 h in the dark at RT; washing with PBS three times. Imaging was performed using a Leica AF6000 microscope. As primary antibodies, chicken anti-GFP (AVES Labs), mouse anti-Ki67 (BD Biosciences) and rabbit anti-active Caspase 3 (R&D Systems) were used. As secondary antibodies, the following were used: CF647-conjugated anti-mouse IgG (Biotium) and CF647-conjugated anti-rabbit IgG (Biotium), FITC-conjugated anti-chick IgY (Biotium). For the EdU assay, cells were cultured with 10 μM EdU in Click-iT EdU Imaging Kits (Life Technologies) for 4 h, fixed with 4% PFA, permeabilized with 0.1% Triton-X and 3% FBS in PBS, and stained with Click-iT reaction buffer for 30 min at RT in the dark. After washing with PBS, primary and secondary staining were performed in the dark.
Cf647 conjugated anti mouse igg
Manufactured by Biotium
CF647-conjugated anti-mouse IgG is a fluorescently labeled secondary antibody that binds to mouse immunoglobulin G (IgG) proteins. The CF647 fluorophore is a far-red dye that can be used for detection and visualization applications.
Automatically generated - may contain errors
Lab products found in correlation
Chicken anti gfp, by Aves Labs (1 mentions)
Rabbit anti active caspase 3, by R&D Systems (1 mentions)
Mouse anti ki67, by BD (1 mentions)
Click it edu imaging kit, by Thermo Fisher Scientific (1 mentions)
Hoechst 33258, by Nacalai Tesque (1 mentions)
Alexa fluor 647 conjugated anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Anti phospho histone h3 ser10, by Merck Group (1 mentions)
Anti e cadherin, by BD (1 mentions)
Anti n cadherin, by BD (1 mentions)
Anti c myc, by Nacalai Tesque (1 mentions)
Anti cux1 sc 13024, by Santa Cruz Biotechnology (1 mentions)
Cf488a anti mouse igg, by Biotium (1 mentions)
Anti zo 1, by Thermo Fisher Scientific (1 mentions)
Anti β catenin, by BD (1 mentions)
Anti tbr1, by Abcam (1 mentions)
Anti nestin, by BD (1 mentions)
2 protocols using cf647 conjugated anti mouse igg
1
Immunocytochemistry Protocol for Cell Imaging
2
Comprehensive Neuronal Cell Marker Profiling
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The following primary antibodies were used for indirect immunofluorescence staining: anti-Cux1 (sc-13024, Santa Cruz Biotechnology); anti-Ctip2 (ab18465, Abcam); anti- GFP (GFP-1010, Aves Labs); anti-Pax6 (PD022, MBL); anti-bromodeoxyuridine (BrdU; catalog #555627, BD Biosciences); anti-phospho-histone-H3 (Ser10; catalog #09-797, Millipore); anti-Rapgef2 (Wei et al., 2007 (link)); anti-Rapgef6 (Yoshikawa et al., 2007 (link)); anti-β-catenin (catalog #610153, BD Biosciences); anti-N-cadherin (catalog #610920, BD Biosciences); anti-E-cadherin (catalog #610181, BD Biosciences); anti-afadin (catalog #ab90809, Abcam); anti-ZO-1 (catalog #61-7300, Life Technologies); anti-nestin (catalog #556309, BD Biosciences); anti-Tbr1 (catalog #ab31940, Abcam); and anti-c-Myc (catalog #23941-54, Nacalai Tesque). To detect immunoreactive signals, the following secondary antibodies were used: Alexa Fluor 488-conjugated anti-chicken Ig Y (catalog #703-545-155, Jackson ImmunoResearch); Alexa Fluor 647-conjugated anti-rabbit IgG (catalog #A21244, Life Technologies); CF647-conjugated anti-mouse IgG (catalog #20281, Biotium); CF488A-conjugated anti-IgG (catalog #20302, #20015, #20027, Biotium); CF488A anti-mouse IgG (Biotium); and CF555-conjugated anti-IgG (catalog #20231, #20038, #20233, Biotium). An Alexa Fluor 647-conjugated anti-Tbr2 antibody (catalog #51-4875, eBioscience) was also used.
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