Axon or soma were lysed in RIPA buffer (Sigma-Aldrich) containing anti-peptidase cocktail (Roche). Lysates were heated in LDS sample buffer and separated on NuPAGE® Novex® 4–12% Bis-Tris Gels (ThermoFisher). Proteins were transferred to a Odyssey nitrocellulose membrane (LI-COR Biosciences), and membranes were blocked with Odyssey blocking buffer (LI-COR Biosciences) for 1 h. Proteins were detected by overnight incubation at 4 ºC with antibodies against TH, 1:1000 (Cell Signaling Technology Cat# 13106, RRID:AB_2798122) or β-actin, 1:1000 (Cell Signaling Technology Cat# 12620, RRID: AB_2797972) in Odyssey blocking buffer containing 0.2% Tween 20 for either 2 h at room temperature or overnight at 4 °C, followed by incubation with goat anti-rabbit IgG (H+L) IRDye 800CW (LI-COR,Lincoln, cat# 926–32211, RRID: AB_621843 at a 1:5,000 dilution in Odyssey blocking buffer containing 0.2% Tween 20 and 0.1% SDS for 1 h at room temperature. The membrane was washed 3 times with Tris-buffered saline/0.1% Tween 20 (TBS–T) between each incubation step. The membrane was then imaged with the Odyssey Infrared Imaging System (LI-COR Biosciences).
Anti peptidase cocktail
Manufactured by Roche
The Anti-peptidase Cocktail is a laboratory reagent designed to inhibit the activity of proteolytic enzymes, known as peptidases, in biological samples. This product helps to prevent the unwanted breakdown of proteins during sample preparation and analysis. The core function of the Anti-peptidase Cocktail is to maintain the integrity of target proteins in the sample, facilitating accurate measurement and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey nitrocellulose membrane, by LI COR (2 mentions)
Nupage novex 4 12 bis tris gel, by Thermo Fisher Scientific (2 mentions)
Odyssey blocking buffer, by LI COR (2 mentions)
Odyssey infrared imaging system, by LI COR (2 mentions)
Ripa buffer, by Merck Group (2 mentions)
Irdye 800cw goat anti rabbit igg h l, by LI COR (1 mentions)
Irdye 800cw conjugated goat anti rabbit igg, by LI COR (1 mentions)
Anti phospho th ser40 antibody, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
2 protocols using anti peptidase cocktail
1
Western Blot Analysis of Protein Expression
2
Immunoblotting of Phosphorylated Tyrosine Hydroxylase
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Axon or soma were lysed in RIPA buffer (Sigma-Aldrich) containing anti-peptidase cocktail (Roche). Lysates were heated in LDS sample buffer and 1 mM dithiothreitol, and separated on NuPAGE Novex 4–12% Bis-Tris Gels (ThermoFisher). Proteins were transferred to a Odyssey nitrocellulose membrane (LI-COR Biosciences), and membranes were blocked with Odyssey blocking buffer (LI-COR Biosciences) for 1 h. Proteins were detected by overnight incubation at 4°C with antibodies against TH (Cell Signaling, dilution 1:1000), anti-phospho-TH (Ser40) antibody (Cell Signaling, dilution 1:500), or β-actin (Cell Signaling, dilution 1:1000) in Odyssey blocking buffer containing 0.2% Tween 20 for either 2 h at RT or overnight at 4°C, followed by incubation with a IRDye 800CW-conjugated goat anti-rabbit IgG (LI-COR Biosciences) at a 1:5000 dilution in Odyssey blocking buffer containing 0.2% Tween 20 and 0.1% SDS for 1 h at RT. The membrane was washed 3 times with Tris-buffered saline/0.1% Tween 20 (TBS–T) between each incubation step. The membrane was then imaged with the Odyssey Infrared Imaging System (LI-COR Biosciences).
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!