Mice were anesthetized with isofluorane and transcardially perfused with cold 0.1M phosphate buffer, followed by 4% paraformaldehyde (PFA) dissolved in 0.1M PB. Brains were post-fixed for 3 hours in 4% PFA at 4˚C and cryoprotected in 30% sucrose. Frozen brains were sliced into 30 μM sections using a cryostat (CM3050, Leica). Free-floating sections from the whole brain were washed 3x in tris-buffered saline (TBS) and then blocked in 1% bovine serum albumin (BSA) in tritonated TBS (0.25% Triton X-100 in TBS) for 1h. The sections were incubated overnight at room temperature in primary antibody (rabbit anti-cFos antibody (Synaptic Systems, #226003, 1:5000), or rabbit anti-SST antibody (Peninsula Laboratories; #T-4103.0050, 1:1000). Slices were then washed in 1% BSA in tritonated BST, followed by incubation with appropriate secondary antibodies (Alex 568 goat anti-rabbit IgG (H+L) (Life Technologies: #A-11036; 1:500), or Alexa 647 goat anti-rabbit IgG (H+L) (Life Technologies: #A-21245, 1:500). Sections were then mounted with DAPI Fluoromount-G (Southern Biotech).
Alexa 647 goat anti rabbit igg h l
Manufactured by Thermo Fisher Scientific
Alexa Fluor 647 goat anti-rabbit IgG (H+L) is a secondary antibody conjugated with the Alexa Fluor 647 fluorescent dye. It is designed to detect and visualize rabbit primary antibodies in various immunoassays and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti c fos antibody, by Synaptic Systems (2 mentions)
Alex 568 goat anti rabbit igg h l, by Thermo Fisher Scientific (2 mentions)
Cm3050, by Leica camera (2 mentions)
Dapi fluoromount g, by Southern Biotech (2 mentions)
Abn78, by Merck Group (1 mentions)
Mowiol 4 88, by Merck Group (1 mentions)
4 protocols using alexa 647 goat anti rabbit igg h l
1
Immunohistochemical Staining of Mouse Brains
2
Immunohistochemical Staining of Mouse Brains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were anesthetized with isofluorane and transcardially perfused with cold 0.1M phosphate buffer, followed by 4% paraformaldehyde (PFA) dissolved in 0.1M PB. Brains were post-fixed for 3 hours in 4% PFA at 4˚C and cryoprotected in 30% sucrose. Frozen brains were sliced into 30 μM sections using a cryostat (CM3050, Leica). Free-floating sections from the whole brain were washed 3x in tris-buffered saline (TBS) and then blocked in 1% bovine serum albumin (BSA) in tritonated TBS (0.25% Triton X-100 in TBS) for 1h. The sections were incubated overnight at room temperature in primary antibody (rabbit anti-cFos antibody (Synaptic Systems, #226003, 1:5000), or rabbit anti-SST antibody (Peninsula Laboratories; #T-4103.0050, 1:1000). Slices were then washed in 1% BSA in tritonated BST, followed by incubation with appropriate secondary antibodies (Alex 568 goat anti-rabbit IgG (H+L) (Life Technologies: #A-11036; 1:500), or Alexa 647 goat anti-rabbit IgG (H+L) (Life Technologies: #A-21245, 1:500). Sections were then mounted with DAPI Fluoromount-G (Southern Biotech).
3
Immunohistochemical Detection of NeuN
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SC slices were incubated in blocking solution (PB 0.1 M, with 4% normal goat antiserum (NGS, Biozol) and 0.4% Triton X-100 (Roth)) for 2 h, then washed 5 times in PB 0.1 M. Subsequently, slices were treated with primary antibody rabbit IgG anti-NeuN (Merck, ABN78) diluted in PB 0.1 M 1:500 with 1% NGS for 48 h at 4°C. Finally, after washing again 5 times with PB 0.1 M, slices were incubated for 2 h at room temperature in secondary antibody goat anti-rabbit IgG (H+L) Alexa 647 (Invitrogen, A32733) diluted 1:500 in PB 0.1 M containing 0.3% Triton X-100 and 3% NGS.
4
Immunofluorescent Labeling of GABA Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SC slices were preincubated in PB 0.1 M containing 1% sodium borohydride for 15 min to remove glutaraldehyde autofluorescence. Subsequently, SC slices were washed 3 times in PB 0.1 M and then incubated in a blocking solution (4% NGS and 0.3% Triton X-100 in PB 0.1 M) for 30 min. We then washed the slices 5 times in PB 0.1 M and incubated them with primary antibody rabbit IgG antiGABA (Sigma A2052) diluted in PB 0.1 M 1:1,000 with 1% NGS overnight at 4°C. The following day, we washed 5 times with PB 0.1 M and incubated them for 1 h at room temperature in secondary antibody goat anti-rabbit IgG (H+L) Alexa 647 (Invitrogen, A32733) diluted 1:1,000 in PB 0.1 M containing 1% NGS. Finally, slices were washed 5 times in PB 0.1 M and mounted with a mounting solution (Mowiol 4–88, 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!