According to the method of our previous studies [28 (link),32 (link)], Western blot analysis was performed. In brief, the ipsilateral dorsal horns of the L5 spinal cord region were collected and used for the experiment. After homogenizing the tissue in PRO-PREPTM (iNtRON Biotechnology, 17081, Kirkland, Washington, USA) and PNPP (Sigma, N9389, St. Louis, MO, USA), lysates were centrifuged. Protein was quantified by Bradford assay (BioRad, # 5000002, Hercules, CA, USA) using only supernatant and 20 μg of protein were used. Proteins were transferred using NC membrane and all blots were blocked with 5% skim milk (BD Biosciences, 90002-594, Franklin Lakes, NJ, USA). p-p66shc (Enzo Life Sciences, ALX-B04-358-C100, 1:1000, Lausen, Switzerland), total shc (BD Biosciences, #6108780, 1:1000), cleaved Caspase-3 (Cell signaling, 9661T, 1:1000), p62 (Sigma, P0067, 1:1000) and PINK1 (Novus Biologicals, NBP2-36488, 1:1000, Centennial, CO, USA) were used as primary antibodies. Visual confirmation of the band was developed using ClarityTM Western ECL Substrate (BioRad, 170-5061).
Nbp2 36488
Manufactured by Novus Biologicals
Sourced in United States
NBP2-36488 is a laboratory equipment product offered by Novus Biologicals. It serves as a tool for researchers and scientists, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Bradford assay, by Bio-Rad (1 mentions)
Pink1, by Novus Biologicals (1 mentions)
Pro prep, by iNtRON Biotechnology (1 mentions)
Clarity western ecl substrate, by Bio-Rad (1 mentions)
Anti lamp1, by Abcam (1 mentions)
Ab128567, by Abcam (1 mentions)
2 protocols using nbp2 36488
1
Western Blot Analysis of Spinal Cord Proteins
2
Indirect and Direct Immunocytochemistry Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For indirect immunocytochemistry (ICC), the following antibodies were used: mouse anti-PINK1 (Novus Biologicals, NBP236488, 1:1000). For direct ICC the following conjugated antibodies were used: Alexa Fluor 647-labelled anti-p-Ser65-Ub (Cell Signalling Technology [CST], 62802, 1:1500), Alexa Fluor 488-labelled anti-HSP60 (Abcam, ab128567, 1:1000), and Alexa Fluor 555 labelled-anti-LAMP1 (BD Biosciences, 555798, 1:1000). Anti-p-Ser65-Ub was conjugated by CST using their basic conjugation service. Anti-HSP60 and anti-LAMP1 were conjugated using Lightning Link Alexa 488 (Abcam, ab236553) and Alexa 555 (Abcam, ab269820) respectively, following the manufacturer's protocol, with an overnight incubation at room temperature.
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!