Mice were perfused with 0.1M PBS and 4% paraformaldehyde and brains were post-fixed for 24 h. For dendritic spine morphology and virus placement, 100 μm sections were blocked in 3% normal donkey serum and 0.3% Triton X-100 in PBS for 30 min, then incubated in chicken anti-mCherry (1:500; Novus Biologicals, NBP2 25158) and in rabbit anti-GFP (1:500; Cell Signaling #2555) overnight at 4°C as in 34 (link), 35 (link). For virus validation a mouse monoclonal anti-Nr4a1 antibody (1:1000; #sc-365113, Santa Cruz) or a polyclonal rabbit anti-SaCas9 (1:1000; #ab203933, Abcam) were used. Following eight 1 h-PBS washes slices were incubated overnight in anti-chicken-Cy3 (1:1000, Jackson Immuno; #703–165-155) and anti-rabbit-Alexa 488 (1:1000, Jackson Immuno; #111–545-003) or anti-mouse Cy2 (1:1000, Jackson Immuno; #715–225-150), anti-rabbit Cy5 (1:1000, Jackson Immuno; #711–175-152) depending on the condition at 4°C. Finally, following eight 1 h-PBS washes, slices were mounted with Aqua-Poly/Mount (Polysciences) mounting media.
Sc 365113
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Sc-365113 is a laboratory product offered by Santa Cruz Biotechnology. It is a piece of equipment designed to assist with scientific research and analysis. The core function of this product is to provide a tool for researchers to conduct their experiments and studies. No further details on the intended use or specific capabilities of Sc-365113 are available at this time.
Automatically generated - may contain errors
Lab products found in correlation
Anti mouse cy2, by Jackson ImmunoResearch (1 mentions)
Anti rabbit cy5, by Jackson ImmunoResearch (1 mentions)
Anti chicken cy3, by Jackson ImmunoResearch (1 mentions)
Aqua poly mount, by Polysciences (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ab7778, by Abcam (1 mentions)
Ab124964, by Abcam (1 mentions)
Ab270993, by Abcam (1 mentions)
Bca assay kit, by Bio-Rad (1 mentions)
Ab9485, by Abcam (1 mentions)
2 protocols using sc 365113
1
Optimized Immunohistochemistry for Dendritic Spines
2
Quantitative Western Blot Analysis of Fibrosis Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The tissue or cell samples were incubated with radioimmunoprecipitation assay (RIPA) protein lysate (Camilo Biological, Nanjing, China) to isolate protein. Protein concentration was detected using the bicinchoninic acid (BCA) assay kit (Bio‐Rad, USA). After denaturation, the protein samples were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and blotted onto polyvinylidene difluoride (PVDF) membranes (Millipore, USA). The membranes were blocked with defatted milk and probed with primary antibodies against PTBP1 (#72669, 1:1000, Cell Signaling Technology, USA), collagen I (ab270993, 1:1000, Abcam, UK), collagen III (ab7778, 1:5000, Abcam), α‐smooth muscle actin (α‐SMA) (ab124964, 1:1000, Abcam), Nur77 (sc‐365113, 1:500, Santa Cruz, USA), FABP5 (#39926, 1:1000, Cell Signaling Technology), and GAPDH (ab9485, 1:2500, Abcam) overnight at 4°C. After incubation with appropriate secondary antibodies (Abcam), the protein bands were detected using an enhanced chemiluminescence (ECL) detection kit (Pierce, USA).
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!