Cells were fixed in 4% paraformaldehyde (PFA) in PBS (w/v) for 10 min, rinsed with PBS, and blocked by 10% normal goat or donkey serum (NGS/NDS) in PBST (PBS + 0.1% Triton X-100) for 60 min. Cells were then incubated with primary antibodies in 10% NGS/NDS in PBST overnight at 4 °C following 1 h of incubation at 24 °C with the appropriate Alexa 488/568 coupled secondary antibodies (1:1000, Invitrogen, MA, USA). Cell nuclei were stained with DAPI; final images were acquired using a Leica TCS SP8 confocal microscope (Leica, Germany) and analyzed using Fiji software. Primary antibodies included rabbit anti-LC3B (1:500, Cell Signaling Technologies #2775, MA, USA), rabbit anti-TBR1 (1:500, Abcam #ab31940, UK), and mouse anti-β-III-tubulin (1:1000, BioLegend, CA, USA. Previously Covance PRB-435P). For live imaging of Aβ in neuronal cultures, cells were incubated with 100 nM BTA-1 and 1 μM NeuroFluor™ NeuO (Invitrogen, MA, USA) for 20 min. The LC3 particle number in β-III-tubulin-positive cells was quantified with the “analyse particles” plug-in in ImageJ (NIH). Quantification was carried out on at least 50 cells per condition, from three independent experiments.
Alexa 488 568 coupled secondary antibodies
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 488/568 coupled secondary antibodies are fluorescent dyes used for detection and visualization in immunoassays. They specifically bind to the Fc region of primary antibodies, allowing for indirect labeling of target proteins or cells. These secondary antibodies provide a bright, stable fluorescent signal for use in techniques such as immunofluorescence microscopy, flow cytometry, and western blotting.
Automatically generated - may contain errors
2 protocols using alexa 488 568 coupled secondary antibodies
1
Immunocytochemistry and Live Imaging of Alzheimer's Markers
2
Immunocytochemistry and Live Imaging of Alzheimer's Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed in 4% paraformaldehyde (PFA) in PBS (w/v) for 10 min, rinsed with PBS, and blocked by 10% normal goat or donkey serum (NGS/NDS) in PBST (PBS + 0.1% Triton X-100) for 60 min. Cells were then incubated with primary antibodies in 10% NGS/NDS in PBST overnight at 4 °C following 1 h of incubation at 24 °C with the appropriate Alexa 488/568 coupled secondary antibodies (1:1000, Invitrogen, MA, USA). Cell nuclei were stained with DAPI; final images were acquired using a Leica TCS SP8 confocal microscope (Leica, Germany) and analyzed using Fiji software. Primary antibodies included rabbit anti-LC3B (1:500, Cell Signaling Technologies #2775, MA, USA), rabbit anti-TBR1 (1:500, Abcam #ab31940, UK), and mouse anti-β-III-tubulin (1:1000, BioLegend, CA, USA. Previously Covance PRB-435P). For live imaging of Aβ in neuronal cultures, cells were incubated with 100 nM BTA-1 and 1 μM NeuroFluor™ NeuO (Invitrogen, MA, USA) for 20 min. The LC3 particle number in β-III-tubulin-positive cells was quantified with the “analyse particles” plug-in in ImageJ (NIH). Quantification was carried out on at least 50 cells per condition, from three independent experiments.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!