Hypothalamus frontal sections of 40 µm were obtained from the rat brains previously fixed by vascular perfusion and post immersion fixation in 4% (w/v) paraformaldehyde for 24 h. Slices were obtained using a vibratome, and processing by free-floating immunohistochemistry. Slices were washed with Tris phosphate buffer (84 mM Na2HPO4, 35 mM KH2PO4, 120 mM NaCl, 10 mM Tris, pH 7.8). BrdU detection was carried out using an additional treatment for allowing DNA denaturation. Samples were incubated in 1 M HCl at 45 °C for 30 min; after that were neutralized by washing the slides three times with Tris phosphate buffer. The following primary and secondary antibodies were used: mouse anti-nestin antibody (1:1500, Abcam Cat# AB6320), chicken anti-vimentin antibody (1:400, Millipore Cat# MAB3400), sheep anti-BrdU antibody (1:1000, Abcam, Cat# AB1893), goat anti-sheep Cy3, donkey anti-chicken Cy2, donkey anti-mouse Alexa 488. All secondary antibodies used were obtained from Jackson Immuno Research and used at a 1:200 dilution. Nuclei was labeled with TOPRO-3 (1:1000, Invitrogen).
Mouse anti nestin antibody
Manufactured by Abcam
Mouse anti-nestin antibody is a primary antibody that specifically binds to the nestin protein, an intermediate filament protein commonly used as a marker for neural stem cells and progenitor cells. This antibody can be used to detect and identify nestin-expressing cells in various applications.
Automatically generated - may contain errors
2 protocols using mouse anti nestin antibody
1
Immunohistochemical Analysis of Rat Hypothalamus
2
Immunofluorescence Analysis of Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
OSCs and differentiated OSCs were fixed in 4% paraformaldehyde (PFA, Wako Pure Chemical Industries, Ltd., Osaka, Japan) for 10 minutes and rinsed 3 times with PBS. Nonspecific binding was blocked with 5% goat serum in PBS for 20 minutes, and spheres and cells were incubated overnight at 4°C with primary antibodies diluted in PBS containing 2% goat serum. The primary antibodies used in this study include mouse anti‐Nestin antibody (Abcam Plc., Cambridge, MA), mouse anti‐Musashi‐1 antibody (Abcam Plc.), mouse anti‐βIII‐tubulin antibody (Abcam Plc.), rabbit antigalactocerebroside (GalC) antibody (Millipore Corp., Billerica, MA), mouse antiglial fibrillary acidic protein (GFAP) antibody (Abcam Plc.), and mouse antiolfactory marker protein (OMP) antibody (Abcam Plc.). Alexa Fluor 488‐ or Alexa Fluor 546‐conjugated goat antirabbit IgG or antimouse IgG antibodies (Life Technologies Corp.) were used as the secondary antibodies. Nuclei were stained with Hoechst 33258 (Dojindo Molecular Technologies, Inc., Kumamoto, Japan) 15 . Images were obtained by confocal laser scanning microscopy (Leica Microsystems GmbH, Wetzlar, Germany). Antibody specificity was confirmed using a universal negative control immunoglobulin (Dako A/S, Glostrup, Denmark).
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!