Kura’s Support Center

• How long will Instant Buffer I last once I dissolve it with dH20?

Instant buffer I is formulated to last for one year upon dilution in LCMS Grade water. However, we recommend to check the buffer for contamination/floating debris on a daily basis. 

• Can I store prepared Instant Buffer I at 2-8 °C?

We recommend storing prepared Instant Buffer I at room temperature, because it can precipitate out of solution when stored in cold temperatures. If refrigerated, make sure crystals are properly solubilized before using it by agitating gently at room temperature until precipitate dissolves. 

• If the enzyme I have in my lab has expired, what are the risks of using it anyway?

The risk is that the enzyme will not have its optimum activity. You can measure activity of the enzyme or contact Kura Biotech at sales@kurabiotech.com to see if a retest of the lot in question is possible. 

• What should I do if my enzyme has been subjected to improper storage conditions?

Please contact Kura Biotech. We can help you determine if the enzyme can be used or needs to be replaced. 

• What are the shelf life and storage conditions of Kura's enzymes?

∙ B-One: 18 months at 2-8°C or 3 months at Room Temperature
∙ BGTurbo: 18 months at 2-8°C
∙ BG100: 12 months at 2-8°C
∙ BGS: 12 months at 2-8°C
∙ ASPC: 12 months at 2-8°C 

• If I'm using BGTurbo, and leave the enzyme over night at room temperature, will this mean I have to discard the enzyme?

BGTurbo glycerol free is stable at room temperature for 14 days, meaning it doesn't lose activity. 

• Can I increase the volume of sample?

Sure! Just be sure to keep a reasonable enzyme:sample ratio for your analyte. For more information please talk with a hydrolysis expert at sales@kurabiotech.com.

• What happens if I use a pH and Temperature outside of the range suggested? 

The enzyme will be working in sub-optimum condition negatively impacting the ability to cleave substrates and you will obtain less recovery. If you need a specific parameter, please contact a hydrolysis expert at sales@kurabiotech.com to help you set up an optimal protocol.

• Does the protocol suggested in the Technical Datasheet of your enzymes apply for any drug class?

No, it doesn't. The protocol of our Technical Datasheets seeks to hydrolyze the hardest-to-cleave analytes, not being necessary to use same enzyme volume, or temperature, or time of incubation for easier to cleave analytes. Please try to find the drug class you will be analyzing in the Applications Page  to find the Application Note for each case.

• May I hydrolyze in the sample preparation columns/plates? (SPE/SLE/Filter)

Assuming the sample prep device has an appropriate hydrophobic frit to hold up the reaction, at temperature, for the entire incubation, yes. The use of a purified enzyme avoids protein precipitation and centrifugation steps, normally necessary when working in presence of high content of protein. Be careful, many SPE or SLE devices will start to flow with gravity at room temperature, while the temperature increases, the flow increases too. 

• How can I ensure I am getting the best performance from my enzyme?

Contant one of our hydrolysis experts at sales@kurabiotech.com or call (800) 332-1448. 

• Sometimes, a B-glucuronidase cannot hydrolyze glucuronides conjugates. Why?

Hydrolysis can be impacted by many variables in protocol or matrix. It is important to optimize protocols for time, temperature, and pH. Considerations need to be made for reaction components such as MeOH, endogenous inhibitors, isomers having steric hindrances as well as non specific binding to surfaces. Kura Hydrolysis experts are ready to help you with specific metabolite and matrix questions. Please reach out to sales@kurabiotech.com or call (800) 332-1448.

• What if I want increased B-One enzyme but not increased dilution of the sample?

The current B-One formulation is proposed for common laboratory practices, if your laboratory has a specific condition that requires more enzyme without further dilution, contact a us at sales@kurabiotech.com or call (800) 332-1448. 

• How can I protect the labile analytes during my hydrolysis?

Labile analytes like morphine can be protected by using prurified enzymes, short incubation times, and mild temperatures.