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发信人: cyberbreeze (风), 信区: Biology
标 题: 另一个western blot问题
发信站: The unknown SPACE (Thu May 23 11:36:07 2002), 转信
After developed with ECL, the signal is pretty strong at first, but faded qu
ickly, then when I try to wash away the substrate and re-develop, no signal
came out! I guess it's because the substrate binding too well with the 2# Ab
-enzyme to be washed off, anyone can solve this problem?
--
※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 140.251.160.69]
发信人: Ras (很颓的猪。), 信区: Biology
标 题: Re: 另一个western blot问题
发信站: The unknown SPACE (Thu May 23 11:51:09 2002), 转信
ft, ECL system the chemifluorescence fades away very quickly.
firt 10sec to a few minutes give the best signal in most casese.
After 30 minutes, 99% you won't pick up any more signal.
【 在 cyberbreeze (风) 的大作中提到: 】
: After developed with ECL, the signal is pretty strong at first, but faded qu
: ickly, then when I try to wash away the substrate and re-develop, no signal
: came out! I guess it's because the substrate binding too well with the 2# Ab
: -enzyme to be washed off, anyone can solve this problem?
--
一年老一年，一日没一日，
一秋又一秋，一辈催一辈，
一聚一离别，一喜一伤悲。
一榻一身卧，一生一梦里。
寻一夥相识，他一会咱一会，都一般相知，
吹一回，唱一回。
※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 152.3.117.238]
发信人: pIgloveEat (猪宝宝), 信区: Biology
标 题: Re: 另一个western blot问题
发信站: The unknown SPACE (Thu May 23 14:35:01 2002) WWW-POST
what does ECL mean?
【 在 Ras (很颓的猪。) 的大作中提到: 】
: ft, ECL system the chemifluorescence fades away very quickly.
: firt 10sec to a few minutes give the best signal in most casese.
:
: After 30 minutes, 99% you won't pick up any more signal.
:
: 【 在 cyberbreeze (风) 的大作中提到: 】
: : After developed with ECL, the signal is pretty strong at first, but faded
qu
: : ickly, then when I try to wash away the substrate and re-develop, no
signal
: : came out! I guess it's because the substrate binding too well with the 2#
Ab
: : -enzyme to be washed off, anyone can solve this problem?
:
:
--
※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 161.130.115.238]
发信人: tlbb (天龙下凡), 信区: Biology
标 题: Re: 另一个western blot问题
发信站: The unknown SPACE (Thu May 23 15:08:04 2002), 站内信件
EVENTUALLY CAN'T LAST...
【 在 pIgloveEat (猪宝宝) 的大作中提到: 】
: what does ECL mean?
: 【 在 Ras (很颓的猪。) 的大作中提到: 】
: : ft, ECL system the chemifluorescence fades away very quickly.
: : firt 10sec to a few minutes give the best signal in most casese.
: : After 30 minutes, 99% you won't pick up any more signal.
: qu
: signal
: Ab
--
※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 129.64.72.228]
发信人: forrestgump (sleepless in Houston), 信区: Biology
标 题: Re: 另一个western blot问题
发信站: The unknown SPACE (Thu May 23 21:12:22 2002) WWW-POST
ECL detection system has a half life of about 30 min (based on the Amersham
kit). however, if you wash away the substrate and develop again, there should
not be much trouble to pick up the signal. although usually the second time
will give you a slightly weaker signal.
what kind of protocol did you follow?
【 在 cyberbreeze (风) 的大作中提到: 】
: After developed with ECL, the signal is pretty strong at first, but faded qu
: ickly, then when I try to wash away the substrate and re-develop, no signal
: came out! I guess it's because the substrate binding too well with the 2# Ab
: -enzyme to be washed off, anyone can solve this problem?
:
--
LIfe is like a box of chocolate.
You never know what is going to be the next.
※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 128.249.148.122]
发信人: special (xxxx), 信区: Biology
标 题: Re: 另一个western blot问题
发信站: The unknown SPACE (Thu May 23 23:35:34 2002) WWW-POST
【 在 cyberbreeze (风) 的大作中提到: 】
: After developed with ECL, the signal is pretty strong at first, but faded qu
: ickly, then when I try to wash away the substrate and re-develop, no signal
: came out! I guess it's because the substrate binding too well with the 2# Ab
: -enzyme to be washed off, anyone can solve this problem?
:
如果用luminescent image analyzer system,it is no problem to record
band,even if it faded so quickly.
--
※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 133.5.48.89]
发信人: forrestgump (sleepless in Houston), 信区: Biology
标 题: Re: 另一个western blot问题
发信站: The unknown SPACE (Fri May 24 12:42:03 2002) WWW-POST
imaging systems are only about 1.5-2 times faster than exposing to X-ray
films, and some of them are actually slower than films.
but ECL's half life is long enough for detection anyway, as long as you do not
wait till it fades away.
【 在 special (xxxx) 的大作中提到: 】
: 【 在 cyberbreeze (风) 的大作中提到: 】
: : After developed with ECL, the signal is pretty strong at first, but faded
qu
: : ickly, then when I try to wash away the substrate and re-develop, no
signal
: : came out! I guess it's because the substrate binding too well with the 2#
Ab
: : -enzyme to be washed off, anyone can solve this problem?
: :
: 如果用luminescent image analyzer system,it is no problem to record
: band,even if it faded so quickly.
:
:
--
LIfe is like a box of chocolate.
You never know what is going to be the next.
※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 128.249.148.55]
发信人: Marble (小石头哥哥), 信区: Biology
标 题: Re: 另一个western blot问题
发信站: The unknown SPACE (Mon May 27 04:41:12 2002), 站内信件
try ECL plus, and avoid exposure your blot to light. do it as quickly
as possible (flurorescence has short halflife). what do you mean by
redevelop? actually i can hardly strip my blots, and there is always
signal remaining.
【 在 cyberbreeze (风) 的大作中提到: 】
: After developed with ECL, the signal is pretty strong at first, but faded qu
: ickly, then when I try to wash away the substrate and re-develop, no signal
: came out! I guess it's because the substrate binding too well with the 2# Ab
: -enzyme to be washed off, anyone can solve this problem?
--
※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 61.157.77.84]
发信人: forrestgump (sleepless in Houston), 信区: Biology
标 题: Re: 另一个western blot问题
发信站: The unknown SPACE (Mon May 27 13:51:49 2002) WWW-POST
I am not sure about ECL plus. but for ECL the 2nd Ab is pretty easy to be
stripped. just follow Amersham's ECL kit's protocol, and I think you should be
able to get rid of all the signals. it always works for me.
【 在 Marble (小石头哥哥) 的大作中提到: 】
: try ECL plus, and avoid exposure your blot to light. do it as quickly
: as possible (flurorescence has short halflife). what do you mean by
: redevelop? actually i can hardly strip my blots, and there is always
: signal remaining.
:
: 【 在 cyberbreeze (风) 的大作中提到: 】
: : After developed with ECL, the signal is pretty strong at first, but faded
qu
: : ickly, then when I try to wash away the substrate and re-develop, no
signal
: : came out! I guess it's because the substrate binding too well with the 2#
Ab
: : -enzyme to be washed off, anyone can solve this problem?
:
:
--
LIfe is like a box of chocolate.
You never know what is going to be the next.
※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 128.249.148.122]
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