Neuromuscular junctions (NMJs) were analyzed in the QL muscle. Muscles from P11 mice from each genotype were immersion fixed in 4% PFA for 20 mins and washed in PBS. Single fibers were teased using fine forceps and washed for 30 mins in PBS supplemented with 0.1M glycine. To stain the postsynaptic part of neuromuscular endplates fibers were incubated with alpha-bungarotoxin-555 antibody for 20 mins and washed in PBS before permeabilization with ice-cold methanol at −20°C for 2 mins. Fibers were then washed in PBS and incubated in a blocking solution containing 10% Donkey Serum in 0.3% PBS-T for an hour before exposure to antibodies against neurofilament - rabbit polyclonal anti-Neurofilament (1:500; Millipore) and synaptophysin - guinea pig polyclonal anti-synaptophysin (1:500; Synaptic Systems, 101 004) at 4°C overnight. Samples were washed in PBS prior to one-hour long incubation with the following secondary antibodies: Alexa Fluor 488 donkey anti-rabbit (Jackson ImmunoResearch, 711–545-152) and Donkey anti-guinea pig-Cy5 IgG (Jackson ImmunoResearch, 706–175-148). Fibers were washed and mounted in 30% glycerol in PBS.
Donkey anti guinea pig cy5 igg
Manufactured by Jackson ImmunoResearch
Donkey anti-guinea pig-Cy5 IgG is a secondary antibody conjugated with the fluorescent dye Cyanine 5 (Cy5). It is designed to detect and visualize guinea pig primary antibodies in various immunoassays and imaging applications.
Automatically generated - may contain errors
2 protocols using donkey anti guinea pig cy5 igg
1
Visualization of Neuromuscular Junctions
2
Visualization of Neuromuscular Junctions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Neuromuscular junctions (NMJs) were analyzed in the QL muscle. Muscles from P11 mice from each genotype were immersion fixed in 4% PFA for 20 mins and washed in PBS. Single fibers were teased using fine forceps and washed for 30 mins in PBS supplemented with 0.1M glycine. To stain the postsynaptic part of neuromuscular endplates fibers were incubated with alpha-bungarotoxin-555 antibody for 20 mins and washed in PBS before permeabilization with ice-cold methanol at −20°C for 2 mins. Fibers were then washed in PBS and incubated in a blocking solution containing 10% Donkey Serum in 0.3% PBS-T for an hour before exposure to antibodies against neurofilament - rabbit polyclonal anti-Neurofilament (1:500; Millipore) and synaptophysin - guinea pig polyclonal anti-synaptophysin (1:500; Synaptic Systems, 101 004) at 4°C overnight. Samples were washed in PBS prior to one-hour long incubation with the following secondary antibodies: Alexa Fluor 488 donkey anti-rabbit (Jackson ImmunoResearch, 711–545-152) and Donkey anti-guinea pig-Cy5 IgG (Jackson ImmunoResearch, 706–175-148). Fibers were washed and mounted in 30% glycerol in PBS.
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!