s*n
2 楼
如果有足够的钱现金买房, 但是偏偏要找lender贷款.
buy up rates and receive high upfront credit, 然后迅速pay off mortgage, 这样
岂不是可以从lender手里赚钱?
银行是通过什么来避免这种现象的?
buy up rates and receive high upfront credit, 然后迅速pay off mortgage, 这样
岂不是可以从lender手里赚钱?
银行是通过什么来避免这种现象的?
x*u
3 楼
GFP表达之后,容易被降解吗?
如果一个蛋白融合GFP之后,蛋白降解是否一定会导致GFP也一起降解了?
如果一个蛋白融合GFP之后,蛋白降解是否一定会导致GFP也一起降解了?
R*a
4 楼
The credit may only be applied to your closing cost. You can't get cash
back, from my understanding.
back, from my understanding.
e*s
6 楼
You did not understand LZ's question. LZ has enough money to pay off the
mortgage. LZ does not need to credit to pay off mortgage. LZ just wants
lower closing cost. I think it is a smart idea, very unconventional (most
people don't have the option :)).
mortgage. LZ does not need to credit to pay off mortgage. LZ just wants
lower closing cost. I think it is a smart idea, very unconventional (most
people don't have the option :)).
x*u
14 楼
谢谢大家的回复。
下面这篇文章是说GFP half life是2.8h吗?
Automated live cell imaging of green fluorescent protein degradation in
individual fibroblasts.
Halter M, Tona A, Bhadriraju K, Plant AL, Elliott JT.
Source
Abstract
To accurately interpret the data from fluorescent proteins as reporters of
gene activation within living cells, it is important to understand the
kinetics of the degradation of the reporter proteins. We examined the
degradation kinetics over a large number (>1,000) of single, living cells
from a clonal population of NIH3T3 fibroblasts that were stably transfected
with a destabilized, enhanced green fluorescent protein (eGFP) reporter
driven by the tenascin-C promoter. Data collection and quantification of the
fluorescence protein within a statistically significant number of
individual cells over long times (14 h) by automated microscopy was
facilitated by culturing cells on micropatterned arrays that confined their
migration and allowed them to be segmented using phase contrast images. To
measure GFP degradation rates unambiguously, protein synthesis was inhibited
with cycloheximide. Results from automated live cell microscopy and image
analysis indicated a wide range of cell-to-cell variability in the GFP
fluorescence within individual cells. Degradation for this reporter was
analyzed as a first order rate process with a degradation half-life of 2.8 h
. We found that GFP degradation rates were independent of the initial
intensity of GFP fluorescence within cells. This result indicates that
higher GFP abundance in some cells is likely due to higher rates of gene
expression, because it is not due to systematically lower rates of protein
degradation. The approach described in this study will assist the
quantification and understanding of gene activity within live cells using
fluorescent protein reporters.
下面这篇文章是说GFP half life是2.8h吗?
Automated live cell imaging of green fluorescent protein degradation in
individual fibroblasts.
Halter M, Tona A, Bhadriraju K, Plant AL, Elliott JT.
Source
Abstract
To accurately interpret the data from fluorescent proteins as reporters of
gene activation within living cells, it is important to understand the
kinetics of the degradation of the reporter proteins. We examined the
degradation kinetics over a large number (>1,000) of single, living cells
from a clonal population of NIH3T3 fibroblasts that were stably transfected
with a destabilized, enhanced green fluorescent protein (eGFP) reporter
driven by the tenascin-C promoter. Data collection and quantification of the
fluorescence protein within a statistically significant number of
individual cells over long times (14 h) by automated microscopy was
facilitated by culturing cells on micropatterned arrays that confined their
migration and allowed them to be segmented using phase contrast images. To
measure GFP degradation rates unambiguously, protein synthesis was inhibited
with cycloheximide. Results from automated live cell microscopy and image
analysis indicated a wide range of cell-to-cell variability in the GFP
fluorescence within individual cells. Degradation for this reporter was
analyzed as a first order rate process with a degradation half-life of 2.8 h
. We found that GFP degradation rates were independent of the initial
intensity of GFP fluorescence within cells. This result indicates that
higher GFP abundance in some cells is likely due to higher rates of gene
expression, because it is not due to systematically lower rates of protein
degradation. The approach described in this study will assist the
quantification and understanding of gene activity within live cells using
fluorescent protein reporters.
i*l
15 楼
i have hands on experience on this, correct, fusion protein with gfp tag is
usually more stable that itself
of course you should confirm the stability when you do subcellular
localization
GFP fuse的蛋白定位 only when the fusion is stable, commensense ba
【在 b******s 的大作中提到】
: 请问这个有很多证据吗?如果这样,那么GFP fuse的蛋白定位岂不是不够准确。很多是
: 蛋白降解后剩下的free GFP?
usually more stable that itself
of course you should confirm the stability when you do subcellular
localization
GFP fuse的蛋白定位 only when the fusion is stable, commensense ba
【在 b******s 的大作中提到】
: 请问这个有很多证据吗?如果这样,那么GFP fuse的蛋白定位岂不是不够准确。很多是
: 蛋白降解后剩下的free GFP?
O*e
16 楼
我比较好奇,你这个加GFP后蛋白变得更稳定的结论是怎么得出来的?测了半衰期了?
考虑了过表达的因素了么?
is
【在 i***l 的大作中提到】
: i have hands on experience on this, correct, fusion protein with gfp tag is
: usually more stable that itself
: of course you should confirm the stability when you do subcellular
: localization
: GFP fuse的蛋白定位 only when the fusion is stable, commensense ba
考虑了过表达的因素了么?
is
【在 i***l 的大作中提到】
: i have hands on experience on this, correct, fusion protein with gfp tag is
: usually more stable that itself
: of course you should confirm the stability when you do subcellular
: localization
: GFP fuse的蛋白定位 only when the fusion is stable, commensense ba
s*y
17 楼
他们用的不是普通的eGFP, 而是destablized eGFP
【在 x****u 的大作中提到】
: 谢谢大家的回复。
: 下面这篇文章是说GFP half life是2.8h吗?
: Automated live cell imaging of green fluorescent protein degradation in
: individual fibroblasts.
: Halter M, Tona A, Bhadriraju K, Plant AL, Elliott JT.
: Source
: Abstract
: To accurately interpret the data from fluorescent proteins as reporters of
: gene activation within living cells, it is important to understand the
: kinetics of the degradation of the reporter proteins. We examined the
【在 x****u 的大作中提到】
: 谢谢大家的回复。
: 下面这篇文章是说GFP half life是2.8h吗?
: Automated live cell imaging of green fluorescent protein degradation in
: individual fibroblasts.
: Halter M, Tona A, Bhadriraju K, Plant AL, Elliott JT.
: Source
: Abstract
: To accurately interpret the data from fluorescent proteins as reporters of
: gene activation within living cells, it is important to understand the
: kinetics of the degradation of the reporter proteins. We examined the
i*l
18 楼
sure, i 'd share. but remember every protein is different
at least it worked for me once.
i was trying to purify a membrane protein, which did not give me full length
on protein gel after purification, and the fusion protein (N term gfp) gave
me at least partial full length fusion, based on the size in sds-page.
every other things were the same, same expression system, same induction
conditions, same extraction method.
but we still could not get enough for crystalization, then gave up, hehe
【在 O******e 的大作中提到】
: 我比较好奇,你这个加GFP后蛋白变得更稳定的结论是怎么得出来的?测了半衰期了?
: 考虑了过表达的因素了么?
:
: is
at least it worked for me once.
i was trying to purify a membrane protein, which did not give me full length
on protein gel after purification, and the fusion protein (N term gfp) gave
me at least partial full length fusion, based on the size in sds-page.
every other things were the same, same expression system, same induction
conditions, same extraction method.
but we still could not get enough for crystalization, then gave up, hehe
【在 O******e 的大作中提到】
: 我比较好奇,你这个加GFP后蛋白变得更稳定的结论是怎么得出来的?测了半衰期了?
: 考虑了过表达的因素了么?
:
: is
相关阅读
中药注射剂应该完全禁用2个相似蛋白,1个SHRNA knockdownRe: nat comm vs pnas为兴趣而工作的日子终于来临!“饿死”癌细胞或许开启未来癌症治疗新篇章Paper help出坑之前的犹豫Re: Paper help海归的好日子到头了中医为什么不担心医疗事故?从长江学者李红良学习在中国看病FBI调查伪造机票及会议开支中国“垃圾论文”数量世界第一,但量变会引起质变的!DNA体积不到细胞核的1%scientist position in biopharma (转载)中南大学2018年国际青年学者“湘江论坛-生命科学论坛”邀请函隔壁大妈又笑了欢天喜地过大年了!首个哺乳动物细胞图谱在浙大诞生哥伦比亚的Tom Jessel 实验室被关门!