如何使用酶的硫酸铵悬浮液?
Working with Enzymes Supplied as Ammonium
Sulfate Suspensions – Technical Note
— If the
ammonium sulfate might interfere with the reaction(s):
1 Remove the enzyme from the refrigerator.
Note: We strongly recommend that ammonium sulfate suspensions not be frozen.
2 Make sure the cap is on tight, and then
invert the vial/bottle several times to provide a milky suspension. This may
take a minute or so. Do not vortex or sonicate; such harsh treatment may
denature some of the enzyme.
3 Using a pipette and sterile wide-bore
pipette tip, remove a portion of the suspension.
4 Transfer to a clean microcentrifuge tube
(or centrifuge tube if working on a larger scale).
5 Pellet the enzyme by centrifuging at
approximately 10,000 – 15,000 x g for 10 minutes. If available, use a
refrigerated microcentrifuge set to a temperature between 2°C and 8°C.
6 Carefully remove as much clear
supernatant as possible; retain. It is not necessary to remove absolutely all
of the ammonium sulfate solution.
7 Dissolve the pellet by adding an
appropriate amount of reaction buffer.
8 Assay the supernatant, and the solution
from the pellet, for protein content and/or enzyme activity.
— If the
ammonium sulfate will not interfere with the reaction(s):
1 As in steps 1-3 above.
2 Add that portion directly to the reaction
buffer.
Note that for an ammonium sulfate
suspension, most of the enzyme will be in solid form. It is likely that only
negligible amounts of enzyme will be in the ammonium sulfate solution.
如何使用酶的硫酸铵悬浮液?
注意:
1 任何的硫酸铵悬浮液的酶都不能冷冻,一定要4-8℃贮存。
2 在硫酸铵的悬浮液中,大部分酶是以固体形式存在的。
一 如果硫酸铵可能会干扰后续的酶反应体系,按照以下程序处理:
1 从4℃中取出酶溶液
2 轻柔缓慢颠倒混匀酶溶液,不要漩涡剧烈震荡溶液
3,4 用无菌吸头吸出适量酶溶液于一灭菌的1.5ml离心管中
5 10000-15000g
4℃离心10分钟
6 尽可能的吸出上清,保留沉淀。不必要彻底去除上清。
7 用适量的酶反应缓冲液溶解沉淀
8 进行后续的酶活分析
二 如果硫酸铵不干扰酶反应,按照以下程序处理:
1 步骤1-4同上
2 直接取适量酶溶液加到酶反应缓冲液中