Western blots were performed for Fc-tagged proteins to confirm expression and secretion of the protein. Protein samples were mixed with 4X SDS with β-mercaptoethanol, boiled for 5 min, and loaded for SDS-PAGE (BioRad) experiments. Proteins were electroblotted onto PVDF membrane (GE Healthcare), blocked with 5% nonfat milk in TBS with 0.1% Tween-20 for 1 h, and incubated overnight at 4 °C with goat anti-human IgG (1:200; Jackson ImmunoResearch Laboratories, Catalog-109-025-003, Lot-117103, 135223). After washing with 1X TBS, the membranes were incubated with mouse anti-goat HRP-conjugated secondary antibody (1:5,000; Santa Cruz Biotechnology, Catalog-sc-2354, Lot-A2017, J1718) for 1 h at room temperature. Membranes were washed in 1X TBS and developed using the ECL Select western blot detection kit (GE Healthcare) for chemiluminescent detection (Omega Lum G). Western blots were performed on samples obtained immediately after media collection and concentration, as well as on protein samples obtained from the Protein G/A beads following aggregation assays.
Ecl select western blot detection kit
Manufactured by GE Healthcare
The ECL Select Western Blot Detection Kit is a chemiluminescent detection system designed for the analysis of proteins in Western blot applications. It utilizes an enhanced chemiluminescent (ECL) substrate to enable the detection of target proteins.
Automatically generated - may contain errors
2 protocols using ecl select western blot detection kit
1
Fc-tagged Protein Expression Validation
2
Protein Expression and Secretion Validation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blots were performed for Fc-tagged proteins to confirm expression and secretion of the protein. Protein samples were mixed with 4X SDS with β-mercaptoethanol, boiled for 5 min, and loaded onto SDS-PAGE gels (BioRad) for electrophoresis. Proteins were electroblotted onto PVDF membrane (GE Healthcare), blocked with 5% nonfat milk in TBS with 0.1% Tween-20 for 1 h, and incubated overnight at 4°C with goat anti-human IgG (1:200; Jackson ImmunoResearch Laboratories, Catalog-109-025-003, Lot-117103, 135223) . After washing with 1X TBS, the membranes were incubated with mouse anti-goat HRP-conjugated secondary antibody (1:5000; Santa Cruz Biotechnology, Catalog-sc-2354, Lot-A2017, J1718) for 1 h at room temperature. Membranes were washed in 1X TBS and developed using the ECL Select Western Blot Detection kit (GE Healthcare) for chemiluminescent detection (Omega Lum G).
Western blots were performed on samples obtained immediately after media collection and concentration as well as protein samples obtained from the Protein G/A beads following aggregation assays.
Western blots were performed on samples obtained immediately after media collection and concentration as well as protein samples obtained from the Protein G/A beads following aggregation assays.
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!