For the incubation experiments, supernatants of cultivations were used. The initial cultivation was a 24 h-cultivation, if not mentioned otherwise, of a single colony in 2 mL of SD2xSCAA media. After the incubation, the culture was harvested by centrifugation at 6000 rpm for 3 min. The supernatant was kept on ice or frozen at − 20 °C. The incubation was performed in 1.5-mL Eppendorf tubes with 200 μL volume of supernatant. As positive controls, standards of purified ZHER3_1-ABD (1.97 mg/mL), ZHER3_1-ZHER3_1-ABD (0.77 mg/mL) and ZHER3_1-ABD-ZHER3_1 (1.34 mg/mL) provided by Affibody AB were used with a concentration of 0.01 g/L or stated otherwise. The experiments with the protease inhibitors were done by adding the protease inhibitor before adding the Affibody standard at the concentration suggested by the supplier. We used the Halt Protease Inhibitor Cocktail EDTA-free (100x) (Thermo Fisher) and 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF) (final concentration 1 mM), aprotinin (final concentration 6.5 μg/mL), bestatin hydrochloride (final concentration 50 µM), leupeptin (final concentration 20 µM), E-64 (final concentration 15 µM), and pepstatin A (final concentration 10 µM). All these chemicals were purchased from Merck.
Bestatin hydrochloride
Manufactured by Thermo Fisher Scientific
Bestatin hydrochloride is a chemical compound used in various research applications. It functions as an inhibitor of aminopeptidase B and leucine aminopeptidase, enzymes involved in protein metabolism. This product is commonly utilized in cell biology, biochemistry, and drug discovery research.
Automatically generated - may contain errors
2 protocols using bestatin hydrochloride
1
Affibody Supernatant Incubation Protocol
2
Protease Inhibitor Evaluation for Protein Stability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the incubation experiments, supernatant of cultivations were used. The initial cultivation was a 24 hcultivation, if not mentioned otherwise, of a single colony in 2 mL of SD2xSCAA media. After the incubation, the culture was harvested by centrifugation at 6000 rpm for 3 min. The supernatant was kept on ice or frozen at -20°C. The incubation was performed in 1.5-mL Eppendorf tubes with 200 µL volume of supernatant. As positive controls, standards of puri ed ZHER3_1-ABD (1.97 mg/mL), ZHER3_1-ZHER3_1-ABD (0.77 mg/mL) and ZHER3_1-ABD-ZHER3_1 (1.34 mg/mL) provided by A body AB were used with a concentration of 0.01 g/L or stated otherwise. The experiments with the protease inhibitors were done by adding the protease inhibitor before adding the A body standard at the concentration suggested by the supplier. We used the Halt Protease Inhibitor Cocktail EDTA-free (100x) (Thermo Fisher) and 4-(2-aminoethyl) benzenesulfonyl uoride hydrochloride (AEBSF) ( nal concentration 1 mM), aprotinin ( nal concentration 6.5 µg/mL), bestatin hydrochloride ( nal concentration 50 µM), leupeptin ( nal concentration 20 µM), E-64 ( nal concentration 15 µM), and pepstatin A ( nal concentration 10 µM). All these chemicals were purchased from Merck.
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!