The regenerated nerve inside the conduit (for the sample withdrawn after 6 weeks) was fixed in 4% paraformaldehyde for 2 h, washed in a solution of 0.01 M PBS (pH 7.2) for 30 min, dehydrated, and embedded in paraffin. Sections were cut 10 μm thick, permeabilized, blocked (0.1% Triton X‐100, 10% normal goat serum for 1 h) and incubated overnight with anti‐NF 200 kDa (monoclonal, mouse, Sigma Aldrich, dilution 1:200) and anti‐S100 (polyclonal, rabbit, Sigma Aldrich, dilution 1:300), at room temperature. Subsequently, sections were washed three times in PBS and incubated for 1 h at room temperature in a solution containing secondary antibodies: Alexa 488 anti‐Mouse (Molecular Probes, dilution 1:200), Cy3 anti‐Rabbit (Life Technologies, dilution 1:300). After three washes in PBS, sections were mounted with a Dako fluorescent mounting and analyzed using a Zeiss LSM800 confocal laser microscopy system (Zeiss, Jena, Germany).
Anti nf 200 kda
Manufactured by Merck Group
The Anti-NF 200 kDa is a laboratory equipment product. It is used to detect and analyze the presence of the 200 kDa neurofilament protein (NF-200) in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti rabbit cy3, by Thermo Fisher Scientific (2 mentions)
Lsm800 confocal laser microscopy system, by Zeiss (2 mentions)
Anti mouse alexa 488, by Thermo Fisher Scientific (2 mentions)
Anti s100, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Fluorescent mounting medium, by Agilent Technologies (1 mentions)
3 protocols using anti nf 200 kda
1
Evaluation of Regenerated Nerve Tissue
2
Immunohistochemical Analysis of Neural Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Crio-embedded longitudinal sections were permeabilized, blocked with 0.1% triton X-100, 10% normal goat serum for 1 h and incubated overnight with the primary antibodies anti-NF 200 kDa (monoclonal, mouse, Sigma Aldrich) and S-100 (polyclonal, rabbit, Sigma Aldrich). After primary antibodies incubation, sections were washed three times in PBS and incubated for 1 h in a solution containing the secondary antibodies Alexa 488 anti-Mouse and Cy3 anti-Rabbit (Life Technologies). Nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI, Sigma) diluted 1:1000 in PBS. After three washes in PBS, sections were mounted with a Dako fluorescent mounting and analyzed using a Zeiss LSM800 confocal laser microscopy system (Zeiss, Jena, Germany).
3
Immunohistochemistry of Axon and Glia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The sections were rinsed in PBS, blocked with normal serum (1% in PBS-Triton 0,1%) for 1 h and then incubated overnight with the primary antibody. After primary antibody(ies) incubation, sections were washed three times in PBS and incubated for 1 h in a solution containing the secondary antibody(ies): Alexa 488 anti-Mouse, Cy3 anti-Rabbit (Life Technologies) conjugated with a fluorophore and selected in order to recognize the species of primary antibodies. After three washes in PBS, sections were finally mounted with a Dako fluorescent mounting medium and stored at 4°C before being analyzed.
Both axon and glia can be detected by immunohistochemistry using specific antibodies. Sections were incubated in a solution containing the following primary antibodies, used alone or in pairs: (i) anti-NF 200 kDa (monoclonal, mouse, Sigma Aldrich); (ii) S-100 (polyclonal, rabbit, Sigma Aldrich).
Both axon and glia can be detected by immunohistochemistry using specific antibodies. Sections were incubated in a solution containing the following primary antibodies, used alone or in pairs: (i) anti-NF 200 kDa (monoclonal, mouse, Sigma Aldrich); (ii) S-100 (polyclonal, rabbit, Sigma Aldrich).
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!