Mice were perfused and brains were postfixed for 24h in 10% formalin. Brain slices were taken using a vibratome (Leica), blocked for 1h with 0.3% Triton X-100, 3% bovine serum albumin (BSA), and 2% normal goat serum (NGS) and incubated in primary antibodies for 24h at 4ºC. Then, free-floating slices were washed three times for 10 min in 0.1% Triton X-100 in PBS (PBS-T), incubated for 1h at room temperature with secondary antibodies, washed in PBS-T and mounted in Vectamount with DAPI (Southern Biotech). Antibodies used here were: anti-cfos (1:500; Cell Signaling Cat#2250S, RRID:AB_2247211), anti-phospho-S6 (Invitrogen, Cat#44–923G, RRID:AB_2533798), anti-mCherry (1:1000; Abcam, Cat# ab205402), anti-GFP (1:1000, Abcam, Cat#ab13970, RRID:AB_300798), anti-Pkcδ (1:500, Abcam, Cat#ab182126) goat-anti-rabbit (Alexa 488 or Alexa 594, Alexa646 1:1000; Thermo Scientific), goat anti-chicken Alexa488, Alexa594, Alexa 647 (1:1000; Thermo Scientific). Images were taken using an LSM780 confocal (Zeiss) and images were processed using ImageJ software (NIH). Cfos counts were conducted for 2–3 sections / animal with a n=2–4 animals / group.
Vectamount with dapi
Manufactured by Southern Biotech
Vectamount with DAPI is a mounting medium formulated for use in fluorescence microscopy. It is designed to preserve fluorescent signals while counterstaining cell nuclei with the DNA-binding dye DAPI.
Automatically generated - may contain errors
Lab products found in correlation
Anti c fos, by Cell Signaling Technology (2 mentions)
Vibratome, by Leica (2 mentions)
Lsm780 confocal, by Zeiss (2 mentions)
Alexa 488, by Thermo Fisher Scientific (2 mentions)
Alexa 647, by Thermo Fisher Scientific (1 mentions)
Alexa 594, by Thermo Fisher Scientific (1 mentions)
Anti mcherry, by Abcam (1 mentions)
Alexa488 goat anti chicken, by Thermo Fisher Scientific (1 mentions)
Axiovert 200 microscope, by Zeiss (1 mentions)
2 protocols using vectamount with dapi
1
Quantifying Neural Activity Markers in Mice
2
Immunohistochemical Analysis of c-Fos Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were perfused, and brains were postfixed for 24 hr in 10% formalin. Brain slices were taken using a vibratome (Leica, Buffalo Grove, IL), blocked for 1 hr with 0.3% Triton X-100, 3% bovine serum albumin (BSA), and 2% normal goat serum (NGS) and incubated in primary antibodies for 24 hr at 4°C. Then, free-floating slices were washed three times for 10 min in 0.1% Triton X-100 in PBS (PBS-T), incubated for 1 hr at room temperature with secondary antibodies, washed in PBS-T and mounted in Vectamount with DAPI (Southern Biotech, Birmingham, AL). Antibodies used here were: anti-c-fos (1:500; Cell Signaling, Danvers, MA), anti-mCherry (1:1000; Abcam, Cambridge, MA) goat-anti-rabbit (Alexa 488 or Alexa 594, 1:1000; Molecular Probes), goat anti-chicken Alexa488 or Alexa594 (1:1000; Molecular Probes). Images were taken using Axiovert 200 microscope (Zeiss, White Plains, NY) or LSM780 confocal (Zeiss) and images were processed using ImageJ software (NIH, Schneider et al., 2012 (link)). C-fos counts were conducted for three or more sections/animal and averaged for each animal for statistical analysis.
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!