Eluted fractions following analytical-SEC were further examined by SDS–PAGE (12% gels) (Bio-rad, California, U.S.A.) and bands were visualised using a Silver staining kit (Invitrogen, California, U.S.A.) for analytical-SEC fraction analysis and Coomassie Brilliant Blue stain (ThermoFisher, Massachusetts, U.S.A.) for band density analysis. Both techniques were performed according to the manufacturer's instructions. Band densities were calculated using GelAnalyzer 9.1 (http://www.gelanalyzer.com/ ).
Sds page 12 gels
Manufactured by Bio-Rad
Sourced in United States
SDS–PAGE (12% gels) is a laboratory equipment used for the separation and analysis of proteins based on their molecular weight. It is a widely used technique in biochemistry and molecular biology for the characterization and identification of proteins.
Automatically generated - may contain errors
Lab products found in correlation
Anti actin antibody, by Santa Cruz Biotechnology (2 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (2 mentions)
Anti il 6, by Novus Biologicals (2 mentions)
Anti tnf α, by Thermo Fisher Scientific (2 mentions)
Nbt bcip, by Thermo Fisher Scientific (2 mentions)
Anti il 1β, by Abcam (2 mentions)
Silver staining kit, by Thermo Fisher Scientific (1 mentions)
Coomassie brilliant blue stain, by Thermo Fisher Scientific (1 mentions)
Anti nf κb, by Thermo Fisher Scientific (1 mentions)
Nf κb, by Thermo Fisher Scientific (1 mentions)
3 protocols using sds page 12 gels
1
Analytical-SEC Protein Fractionation and Visualization
2
Western Blot Analysis of Inflammatory Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed according to a standard protocol [34 (link),35 (link)]. Equal amounts of protein (40 μg/well) were subjected to SDS-PAGE (12% gels, Bio-Rad, Hercules, CA, USA) and transferred to nitrocellulose membranes. After blocking in TBST (Tris-buffered saline, 0.1% Tween 20) with 5% BSA (Bovine Serum Albumin, Invitrogen, Waltham, MA, USA), membranes were probed overnight at 4 °C with the corresponding primary antibodies, e.g., anti-IL-1β (1:500; Abcam, Cambridge, UK); anti-IL-6 (1:750; Novus Biologicals, Centennial, CO, USA); anti-TNF-α, (1:1000; Invitrogen, Waltham, MA, USA); anti-NF-κB, (1:1000; Invitrogen, Waltham, MA, USA). Anti-actin antibody (1:1000, Santa Cruz, CA, USA) was used for the loading control. After the washing procedure, membranes were incubated with alkaline phosphatase-conjugated secondary antibodies, IgG (1:5000 Santa Cruz, CA, USA) for 1 h at room temperature. The immunoreactive bands were visualized by a colorimetric detection kit (NBT-BCIP; ThermoFisher, Waltham, MA, USA) and protein amounts were analyzed with the ImageJ program (1.46r, NIH, Bethesda, MD, USA).
3
Quantitative Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was performed according to a standard protocol [34, 35] . Equal amounts of protein (40 μg/well) were subjected to SDS-PAGE (% 12 gels, Biorad,USA) and transferred to nitrocellulose membranes. After blocking in TBST (Tris-buffered saline, 0.1% Tween 20) with 5% BSA (Bovine Serum Albumin-Invitrogen,USA), membranes were probed overnight at 4°C with the corresponding primary antibodies, e.g. anti-IL-1β (1:500; Abcam,UK); anti-IL-6 (1:750; Novus,USA); anti-TNF-α, (1:1000; Invitrogen,USA) ; NF-κB, (1:1000; Invitrogen,USA). Anti-actin antibody (1:1000, Santa Cruz,USA) was used for the loading control. After the washing procedure, membranes were incubated with alkaline phosphatase conjugated secondary antibodies, IgG (1:5000 Santa Cruz, USA) for 1 hour at room temperature. The immunoreactive bands were visualized by a colorimetric detection kit (NBT-BCIP; ThermoFisher, USA) and protein amounts were analysed with ImageJ programme (1.46r, NIH 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!