in western for a 25kd protein, the band is at 50kd# Biology - 生物学
n*l
1 楼
m*p
2 楼
western for a 25kd protein. using serum.
but the band is clearly at 50kd.
seems dimer, but in sds-page, proteins are denatured, so should not form
dimmer. any suggestions?
thank you very much.
but the band is clearly at 50kd.
seems dimer, but in sds-page, proteins are denatured, so should not form
dimmer. any suggestions?
thank you very much.
s*c
3 楼
grocery, supermarket
C*e
4 楼
加urea呢
T*t
5 楼
To be eligible for this offer, you must be the named recipient of this
invitation. 是不是没收到invitation的就不能注册？
invitation. 是不是没收到invitation的就不能注册？
q*g
6 楼
protein modification?
j*o
7 楼
是的.这是TARGETED OFFER
【在 T**********t 的大作中提到】
: To be eligible for this offer, you must be the named recipient of this
: invitation. 是不是没收到invitation的就不能注册？
S*s
8 楼
60KD nrf2 shows up in 110KD in SDS PAGE.
nobody know the reason for many years.
所以可能性很多
【在 m******p 的大作中提到】
: western for a 25kd protein. using serum.
: but the band is clearly at 50kd.
: seems dimer, but in sds-page, proteins are denatured, so should not form
: dimmer. any suggestions?
: thank you very much.
nobody know the reason for many years.
所以可能性很多
【在 m******p 的大作中提到】
: western for a 25kd protein. using serum.
: but the band is clearly at 50kd.
: seems dimer, but in sds-page, proteins are denatured, so should not form
: dimmer. any suggestions?
: thank you very much.
a*n
9 楼
for some dimers, SDS is not enough to fully disrupt it.
【在 m******p 的大作中提到】
: western for a 25kd protein. using serum.
: but the band is clearly at 50kd.
: seems dimer, but in sds-page, proteins are denatured, so should not form
: dimmer. any suggestions?
: thank you very much.
【在 m******p 的大作中提到】
: western for a 25kd protein. using serum.
: but the band is clearly at 50kd.
: seems dimer, but in sds-page, proteins are denatured, so should not form
: dimmer. any suggestions?
: thank you very much.
t*y
10 楼
”using serum“.会不会是因为血清里的IgG Heavy Chain 污染？
b*y
11 楼
nod. I have many such examples. For instance, I have a 14mer of 10 KDa
protein. But its molecular weight is about 140 KDa on SDS-PAGE.
【在 a*********n 的大作中提到】
: for some dimers, SDS is not enough to fully disrupt it.
protein. But its molecular weight is about 140 KDa on SDS-PAGE.
【在 a*********n 的大作中提到】
: for some dimers, SDS is not enough to fully disrupt it.
u*d
12 楼
这么神奇？什么蛋白？
【在 b******y 的大作中提到】
: nod. I have many such examples. For instance, I have a 14mer of 10 KDa
: protein. But its molecular weight is about 140 KDa on SDS-PAGE.
【在 b******y 的大作中提到】
: nod. I have many such examples. For instance, I have a 14mer of 10 KDa
: protein. But its molecular weight is about 140 KDa on SDS-PAGE.
D*9
13 楼
add 50-100 mM DTT in your loading buffer
k*0
14 楼
样品加高浓度的DTT或TCEP没有？
b*y
15 楼
It is a yeast protein over-expressed in E. coli, in which its authentic
binding partner doesn't exist.
【在 u**********d 的大作中提到】
: 这么神奇？什么蛋白？
binding partner doesn't exist.
【在 u**********d 的大作中提到】
: 这么神奇？什么蛋白？
s*e
16 楼
最大的可能性就是个SDS抗性的二聚体。
SDS-resistant的蛋白多聚体挺多的，老麦的KcsA就是个例子。
SDS-resistant的蛋白多聚体挺多的，老麦的KcsA就是个例子。
m*p
17 楼
have not tried urea. will try it.
have tried increasing dtt, but not that high. will increase even high.
modification is possible. will try a kit that removes glycosilation.
thanks guys. hopefully will update you with new results.
have tried increasing dtt, but not that high. will increase even high.
modification is possible. will try a kit that removes glycosilation.
thanks guys. hopefully will update you with new results.
a*a
18 楼
可能有修饰，也可能是蛋白自身交联
前阵子science上的jmjd6就是自身交联，大小是理论值的2倍 sds page破不开
前阵子science上的jmjd6就是自身交联，大小是理论值的2倍 sds page破不开
g*5
19 楼
68kDa.
and sometime 100kDa, sometime 110kDa.
I am crazy with this one.
and most commercial antibodies do not work...
do you have a good one for mouse tissue?
(no H300)
【在 S*********s 的大作中提到】
: 60KD nrf2 shows up in 110KD in SDS PAGE.
: nobody know the reason for many years.
: 所以可能性很多
and sometime 100kDa, sometime 110kDa.
I am crazy with this one.
and most commercial antibodies do not work...
do you have a good one for mouse tissue?
(no H300)
【在 S*********s 的大作中提到】
: 60KD nrf2 shows up in 110KD in SDS PAGE.
: nobody know the reason for many years.
: 所以可能性很多
S*s
20 楼
Because in the past 10 years, people thought that 68KD is the right one.
H300 works at 1:500 to 1:1000.
That's the only one worked in my hand.
【在 g*********5 的大作中提到】
: 68kDa.
: and sometime 100kDa, sometime 110kDa.
: I am crazy with this one.
: and most commercial antibodies do not work...
: do you have a good one for mouse tissue?
: (no H300)
H300 works at 1:500 to 1:1000.
That's the only one worked in my hand.
【在 g*********5 的大作中提到】
: 68kDa.
: and sometime 100kDa, sometime 110kDa.
: I am crazy with this one.
: and most commercial antibodies do not work...
: do you have a good one for mouse tissue?
: (no H300)
m*p
21 楼
then, how did people know the band at 110kd is jmjd6?
H*g
22 楼
How did u boil the sample before loading on SDS-PAGE?
Do u think if this step could affect the molecular weight of the observed
band(s)?
Do u think if this step could affect the molecular weight of the observed
band(s)?
m*p
23 楼
i boiled it for 5 min.
a website says no boiling can remove dimer, so i tried no boilding, but
still the same result.
a website says no boiling can remove dimer, so i tried no boilding, but
still the same result.
C*I
24 楼
有时SDS不足以解开oligomers。多加些SDS(1.5%)和beta-mercaptoethanol（4%）。然
后在90C加热10分钟试试。
【在 m******p 的大作中提到】
: western for a 25kd protein. using serum.
: but the band is clearly at 50kd.
: seems dimer, but in sds-page, proteins are denatured, so should not form
: dimmer. any suggestions?
: thank you very much.
后在90C加热10分钟试试。
【在 m******p 的大作中提到】
: western for a 25kd protein. using serum.
: but the band is clearly at 50kd.
: seems dimer, but in sds-page, proteins are denatured, so should not form
: dimmer. any suggestions?
: thank you very much.
f*9
25 楼
How about -S-S- linked dimer?
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