Cortical neurons from embryonic day 18 (E18) Sprague Dawley rats were plated at a density of 8.5 × 105 cells per well onto 6-well plates previously coated with poly-d -lysine. The cells were cultured in Neurobasal media containing 0.5 mM L-glutamine and B27 supplement (Life Technologies, Carlsbad, CA, USA) at 37 °C in a humidified atmosphere of a 5% CO2 incubator. Every three days, a half exchange was performed on the medium. After 14 days in vitro (DIV 14), the cells were treated with 100 mM METH and 100 mM oxy for 24 h, following which the cells were collected in ice-cold PBS, and proteins were isolated using an NPer kit (ThermoFisher, Waltham, MA, USA).
Nper kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The NPer kit is a laboratory equipment designed for protein purification. It utilizes a specific technique to isolate and concentrate target proteins from complex samples. The core function of the NPer kit is to enable efficient protein extraction and purification for further analysis or research applications.
Automatically generated - may contain errors
Lab products found in correlation
B27 supplement, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Dctm protein assay, by Bio-Rad (1 mentions)
Anti flag m2 mab, by Merck Group (1 mentions)
Mouse anti β actin c4, by Santa Cruz Biotechnology (1 mentions)
Goat anti lamin b c 20, by Santa Cruz Biotechnology (1 mentions)
3 protocols using nper kit
1
Isolation of Proteins from Methamphetamine and Oxycodone-Treated Cortical Neurons
2
Cell Lysis and Protein Extraction Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in RIPA buffer [50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 1% (v/v) NP-40, 0.5% (w/v) deoxycholate, and 0.1% (w/v) SDS, supplemented with phenylmethylsulfonyl fluoride, sodium orthovanadate, and a cocktail of protease inhibitors. The N-Per kit (Thermo Fisher Scientific) was used for cellular fractionation. Protein concentrations of the cell lysates were measured by the DCTM Protein assay (Bio-Rad Laboratories).
3
Murine T Cell Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole-cell or nuclear extracts from primary murine T cells were prepared with the M- or N-PER Kit (Thermo Fisher Scientific), respectively. Proteins were resolved by 8–12% SDS-PAGE followed by immunoblotting. The primary antibodies used were rat anti-Bcl2A1 (#11470; Lang et al., 2014 (link)), mouse anti-NFATc1 (7A6; BD PharMingen), rabbit anti-NFATc2 (IG-209; ImmunoGlobe), and goat anti–Blimp-1 (C-21), mouse anti–β-actin (C-4), goat anti-lamin B (C-20), and anti-ERK (H-72; all Santa Cruz Biotechnology) with anti-mouse, -rabbit, or -goat peroxidase-coupled secondary antibodies. 3 µg of nuclear extracts from EL-4 cells or pEYZ/Blimp-1F (Schmidt et al., 2008 (link))-transfected HEK 293T cells were prepared and subjected to EMSA as described (Berberich-Siebelt et al., 2000 (link), 2006 (link)). Antibodies for supershifts were anti-NFATc1 (AB1–205; ImmunoGlobe), anti-NFATc2 and –Blimp as above, and anti-Flag mAb (M2; 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!