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Would protease inhibitor cocktail in my protein buffer interfere...
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Masao Miwa
Would protease inhibitor cocktail in my protein buffer interfere...
Some of the protease inhibitors are short peptides and small proteins, so they will give a signal in the protein assay. If they remain in the sample, then you can compensate for that by including the same concentration of protease inhibitors in the standards. However, the interference from the protease inhibitors may be negligible of your sample is reasonably concentrated.
A problem with bead beating is the amount of heat that is generated, which will accelerate the activity of proteolytic enzymes. Do your best to keep the sample cold by using short bursts of bead beating interspersed with cooling the sample in an ice water bath.
Some of the protease inhibitors are short peptides and small proteins, so they will give a signal in the protein assay. If they remain in the sample, then you can compensate for that by including the same concentration of protease inhibitors in the standards. However, the interference from the protease inhibitors may be negligible of your sample is reasonably concentrated.
A problem with bead beating is the amount of heat that is generated, which will accelerate the activity of proteolytic enzymes. Do your best to keep the sample cold by using short bursts of bead beating interspersed with cooling the sample in an ice water bath.
Many protease inhibitors are highly reactive chemicals (PMSF, DTT) and could interfere. The vendor should have info although they can't know every system. Luckily these are easily done control experiments, maybe worthwhile to do? Good luck!
Many protease inhibitors are highly reactive chemicals (PMSF, DTT) and could interfere. The vendor should have info although they can't know every system. Luckily these are easily done control experiments, maybe worthwhile to do? Good luck!
We have specifically used HALT and have had no interference in our BCA assay from biorad. You could always add a control of lysis buffer alone with protease inhibitor if you want to negate the effects.
We have specifically used HALT and have had no interference in our BCA assay from biorad. You could always add a control of lysis buffer alone with protease inhibitor if you want to negate the effects.
Some of the protease inhibitors are short peptides and small proteins, so they will give a signal in the protein assay. If they remain in the sample, then you can compensate for that by including the same concentration of protease inhibitors in the standards. However, the interference from the protease inhibitors may be negligible of your sample is reasonably concentrated.
A problem with bead beating is the amount of heat that is generated, which will accelerate the activity of proteolytic enzymes. Do your best to keep the sample cold by using short bursts of bead beating interspersed with cooling the sample in an ice water bath.
Some of the protease inhibitors are short peptides and small proteins, so they will give a signal in the protein assay. If they remain in the sample, then you can compensate for that by including the same concentration of protease inhibitors in the standards. However, the interference from the protease inhibitors may be negligible of your sample is reasonably concentrated.
A problem with bead beating is the amount of heat that is generated, which will accelerate the activity of proteolytic enzymes. Do your best to keep the sample cold by using short bursts of bead beating interspersed with cooling the sample in an ice water bath.
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Many protease inhibitors are highly reactive chemicals (PMSF, DTT) and could interfere. The vendor should have info although they can't know every system. Luckily these are easily done control experiments, maybe worthwhile to do? Good luck!
Many protease inhibitors are highly reactive chemicals (PMSF, DTT) and could interfere. The vendor should have info although they can't know every system. Luckily these are easily done control experiments, maybe worthwhile to do? Good luck!
More
VOTE
We have specifically used HALT and have had no interference in our BCA assay from biorad. You could always add a control of lysis buffer alone with protease inhibitor if you want to negate the effects.
We have specifically used HALT and have had no interference in our BCA assay from biorad. You could always add a control of lysis buffer alone with protease inhibitor if you want to negate the effects.
More
VOTE