O***n
发帖数: 13127 | 1 要大剂量。所以要吃2 big tablespoons
people told me this. the acid is to use to denature the pollen? haha, not
really sure.
as an real immunologist, I doubt the honey will really work. but no harm
trying it.
the purpose is to induce immunological tolerance through mucosal immune
system. but your allergy occurs first, so it is difficult to induce
tolerance later |
|
B******1
发帖数: 9094 | 2 But your lab probably has proteins, even plant proteins, which could be
denatured to make tofu. Heck if you want to synthesize a unique plant
protein with an iBull signature built-in, you might be able to do it
yourself using a peptide synthesizer. Then you can make a nano-size mini-
protein and publish the result in nature or science as the first step
towards personalize protein for mass consumption.
Mention my name in the acknowledge section in the end, please. |
|
U***J
发帖数: 5998 | 3 Those 200 proof EtOH should be denatured to prevent one from drinking it,
and should not have MeOH though.
stores
a
bottle,
distilled
for |
|
U***J
发帖数: 5998 | 4 My bad, just realized MeOH is one of the additives used to denature EtOH.
Darn it. |
|
m***o
发帖数: 17656 | 5 【 以下文字转载自 Shaanxi 讨论区 】
发信人: miluo (twitter), 信区: Shaanxi
标 题: 烤火鸡之化学研究/详细做法/实战结果
发信站: BBS 未名空间站 (Sun Nov 27 15:55:48 2011, 美东)
和以往一样，在做大餐前，要理论研究一番。
去年做的烤火鸡，按照网上的做法，用apple cider 浸泡，竟被美食家鄙视：不就是苹
果的味道吗？今年上网研究一下，apple cider 和 apple juice 究竟是什么？有什么区
别，这是搜到的：
Apple juice and apple cider are both fruit beverages made from apples, but t
here is a difference between the two. Fresh cider is raw apple juice that ha
s not undergone a filtration process to remove coarse particles of pulp or s
ediment. It t... 阅读全帖 |
|
K****N
发帖数: 10783 | 6 转载
My Top 21 Favorite Foods to Mix with Protein Powder
1. Milk - Just use milk instead of water for more flavor, more thickness,
and more calories. Try goat’s milk for an extra health kick.
2. Cream - Use instead of milk or water for way more flavor and calories.
3. Fruit - Blend it up and it’s called a fruit smoothie. Try
strawberries and bananas. Add fruit to just about all of these other recipes
. Fruit adds vitamins, minerals, and antioxidants so don’t hesitate to add
it to anything... 阅读全帖 |
|
v******s
发帖数: 6949 | 7 fat free milk = sugar water + denatured protein. You are better off drinking
蛋花汤。 |
|
i*********s
发帖数: 8706 | 8 两个产品大部分成分一样。。。
Water
Bifida Ferment Lysate
Glycerin
Alcohol Denatured
Dimethicone
Hydroxyethylpiperazine Ethane Sulfonic Acid
PEG-20 Methyl Glucose Sesquistearate
Sodium Hyaluronate
Ammonium Polyacrylatedimethyl Taurate
Disodium EDTA
Caprylyl Glycol
Citric Acid
Xanthan Gum
Octyldodecanol
Limonene
Fragrance
在此之上，兰蔻黑瓶含有：
Phenoxyethanol。。。。。。。。。。一种preservative
PEG-60 Hydrogenated Castor Oil。。不懂
Salicyloyl Phytosphingosine 。。。不懂
Citronellol 。。。。。。。。。。。不懂
欧莱雅含有：
Palmitoyl Oligopeptide 。。。防老
Palmitoyl... 阅读全帖 |
|
b*s
发帖数: 82482 | 9 应该是白喝了……
The body is very sensitive to its pH level, so strong mechanisms exist to
maintain it. Outside the acceptable range of pH, proteins are denatured and
digested, enzymes lose their ability to function, and death may occur.
能变一点吧。 要不我的健康醋白喝了
呢。 |
|
|
y******e
发帖数: 5906 | 11 如果鸡蛋白因为和铜离子结合变性了，我总觉得重金属离子有毒，还是少这样干好一点。
我一般用不锈钢碗打发蛋白。
再说打发蛋白，是个物理过程，主要是填充空气进去，和化学反应基本无关，所以用不
用铜器都无所谓。
denature |
|
T*******I
发帖数: 5138 | 12 Home Depot有一种Denatured Alcohol。是一种纯酒精（纯度应该高达99.9%以上）。
我想现在一大桶也就15刀以内。十年前我买过一桶，当时价格为10美刀。 |
|
r****r
发帖数: 1839 | 13 不是学生物或化学的吧，洗涤剂把病毒都denature失去活性了。 |
|
g******i
发帖数: 581 | 14 杨振宁 国立西南联合大学-学士(1942),硕士(1944)
1957年以中华民国公民身份获得诺贝尔物理学奖，1986年获美国国家科学奖章，1993年
获本杰明.富兰克林奖章，1995年获 爱因斯坦奖章，与李政道提出弱相互作用中宇称不
守恒.与罗伯特·米尔斯一道提出了杨-米尔斯理论，即非阿贝尔规范理论，对基础物理
学产生了深远的影响，是粒子物理学的标准模型的基础
李政道 国立浙江大学物理系/国立西南联合大学-学士 芝加哥大学博士
1957年以中华民国公民身份与杨振宁以弱作用下宇称不守恒的的发现获得诺贝尔物理学奖
吴健雄 国立中央大学数学/物理学士，先后在国立浙江大学,中央研究院物理研究所工
作美国国家科学院院士 美国国家科学奖章获得者，沃尔夫奖获得者，曾任美国物理学
会会长
1957年验证杨振宁李政道的“弱相互作用下的宇称不守恒”，1963年实验证明“β 衰
变在矢量流守恒定律”
在制造原子弹的“曼哈顿计划”中解决了链式反应无法延续的重大难题
被美国物理学会宣布为最伟大的实验物理学家之一
Madam Wu is arguably the most admired female sci... 阅读全帖 |
|
|
T******e
发帖数: 18290 | 16 美国国籍有这么容易放弃？一般只有成年人做出有放弃美国国际intention的事情，才
会被认为放弃美国国籍。这个小孩生在美国，甚至不是归化公民，不可能因为其父母的
决定丧失美国国籍。
With a few exceptions, anyone with U.S. citizenship will retain it for life.
The exceptions include when one of the following takes place:
The U.S. immigration authorities revoke the person’s naturalized
citizenship. Called “denaturalization,” this will happen only if you
obtained your citizenship illegally in the first place, through fraud or
concealment of a material fact, or willful misrepresentation.
... 阅读全帖 |
|
|
h**l
发帖数: 4883 | 18 Do an experiment. Put a slice of chicken in the oven at 133 F for 2 days and
see if the meat is raw or not.
I think most of the meat protein will denature at that environment. But that
temperature can't kill pathogens. Lack of humidity might kill some of the
pathogens. |
|
m***j
发帖数: 9290 | 19 cooked/raw and denature are different concepts.
and
that |
|
x*****a
发帖数: 972 | 20 家里发现有pill bug（西瓜虫/潮虫），虽然不咬人什么的，但是看着烦。而且很多聚
集在heating system的compartment里，无数的尸体，因为我家其实比较干，我把暖气开到最大
了。我用杀虫剂喷了下，感觉效果不好，因为那杀虫剂是针对蟑螂的，但按理说，既然可以denature
protein，应该啥都能杀了吧。
我google了下，发现多数的杀虫剂都是针对户外草坪的，我需要个室内的。大家有什么好办法么？房东
不是很有用，而且我有猫，我白天都不在家，不想让外人进我家，我猫非常怕陌生人，特别是在我不在
场的情况下。 |
|
c****y
发帖数: 584 | 21 homedepot有号称organic的杀虫剂， 去看看？
气开到最大
然可以denature
么好办法么？房东
，特别是在我不在 |
|
|
h******n
发帖数: 493 | 23 The sensitivity of detecting the intact protein
complexes by MS directly is still not very good,
not as good as detecing just proteins under the
denatured conditions.
But people usually immunoprecipitate the protein
complexes and then run gels or use other methods
to separate the proteins, and then identify the
ligand proteins by MS. This method is very sensitive
and accurate.
Hope this answer your question. |
|
m***o
发帖数: 17656 | 24 和以往一样，在做大餐前，要理论研究一番。
去年做的烤火鸡，按照网上的做法，用apple cider 浸泡，竟被美食家鄙视：不就是苹
果的味道吗？今年上网研究一下，apple cider 和 apple juice 究竟是什么？有什么区
别，这是搜到的：
Apple juice and apple cider are both fruit beverages made from apples, but t
here is a difference between the two. Fresh cider is raw apple juice that ha
s not undergone a filtration process to remove coarse particles of pulp or s
ediment. It takes about one third of a bushel to make a gallon of cider.
Apple juice is juice that has been filtered to remove solids and past... 阅读全帖 |
|
c*******r
发帖数: 6 | 25 there are at least 12 caspases rigt now, they are all cystein proteases,
cut after asp (D) thus the name caspases. they are homologous to CED-3.
they can be divided into different groups, seems caspase-1 (ICE interleukin -1
converting enzyme) and some othe caspases are not very important in apoptosis,
but may play a role in immune responce. the other caspases are important in
apoptosis. they can be further divided into several groups by their role in
apoptosis, caspase 8 and 9 are thought to be |
|
m****g
发帖数: 13 | 26 1. buy enough amount of commercially available herring (or salmon) sperm DNA.
They are dirty cheap. you can get 100mg/$200 probably anywhere.
2. digest your DNA using AluI restriction enzyme. If AluI does not work
well, try other enzymes that recognize 4-nucleotide palingdromic sequence.
You may combine 2 enzymes in one digestion mixture.
3. run a 6% denatured sequencing gel using a thicker spacer set (like 1mm), or
better (maybe) a 1.5% agarose gel to seperate your DNA.
4. Cut out y |
|
r****o
发帖数: 105 | 27 Do you have a way to distinguish correctly folded protein? If you have,
just use HPLC to purify them from crude E.Coli expressed protein.
If you don't have one way to do that.
Refolding involves (1) denaturing and reducing (2) suddently dilute
the sample extensively to mild condition (3) get the soluble protein.
Try to search pubmed for methods or refer to any classic protein biochemistry
protocol book. Note: the yield is low(less than 10% of the crude protein
will be correctly refolded). |
|
M****e
发帖数: 70 | 28 my point is to make sure that you have transfer of good
quality and quantity of RNA onto the blotting material.
the washing stringency for your experiment is not quite
high (i.e., 42C). i think the problem is mainly due to
bad RNA or probe. if there is a lot of radioactive
nucleotides incorporated into the probe, you should
detect very hot signal after purification. denature the
probe before use. GAPDH is a housekeeping gene that is
typically used for normalization of load. i suggest you
read a |
|
r****o
发帖数: 105 | 29 It is OK! Actually that's the whole basis for FACS analysis.
The point of fixation is not to denature the protein. It just
crosslinks the proteins and make the organelle structures stable in
a cell.
The antibodies used in IF are usually raised against antigen
in its native structure. |
|
f*********p
发帖数: 13 | 30 desalting column is much faster, but sometimes proteins bind to it
non-specifically and you may lose some of your sample by that. sometimes,
proteins get denatured because the buffer change is too sudden, not like
dialysis that is slow and "tender". plus, carry over is almost inevitable on
the column, and a 2nd run is sometimes necessary.
dialysis on the other hand, has its disadvantage too. most of all, it takes
way too long (O/N is routine), during which the protein may not be happy.
second, i |
|
n********k
发帖数: 2818 | 31 I used primestar from Takara, it is great with my experience...no denaturing
reagents though, otherwise any
Polymerases(I have ever used) would do poorly... |
|
p**********t
发帖数: 2636 | 32 PrimeStar is a really sensitive enzyme. It fails my PCR a lot of the time...
Don't know why. BTW I use Takara ExTaq for my regular PCR, using shorter
priming time for PrimeStar then it stops working.
denaturing |
|
n********k
发帖数: 2818 | 33 google is ur best friend...anyway, in general, it is denature reagent and/or
detergent including DMSO, Betaine, NP40, Triton-X100 etc etc...what I heard
Betaine is likely the best for most of application...and u can buy it from
some company...nonetheless, it could still be tricky even if you know the
gradient, it could still take a hell a lot effort to figure out correct
composition...I personally have used mostly DMSO, and Q solution from qiagen
and a bit betaine... |
|
g*****3
发帖数: 107 | 34 最近在提纯一个GST-tag的蛋白，蛋白表达在pellet需要denature condition提纯， 纯
化后需要做活性检测。怎样才能让蛋白的活性在提纯过程中不丢失呢，需要用
refolding kit吗？谢谢 |
|
f*********r
发帖数: 1233 | 35 与细胞/组织匀浆不同，血清中dna极低。粘稠主要因为蛋白和血脂。楼上说超高速离心
，是个不错的办法。能够去掉很多悬浊微粒。如果样品量有限，可以加过lysis buffer
以后再离心。
稀释总蛋白浓度。如果5ul/well这个量不能再降低，可以象楼上所说，用大胶、厚胶，
想办法增加volume/well。
如果有条件，可以用filter过滤掉不需要的蛋白。一般，条带扭曲多发生在小分子量。我估计你要的蛋白就不大。那么，你可以根据分子量，过滤掉大分子蛋白。millipore就有很多不同孔径的过滤系统可供选择。
条带形状不规整，很大程度上还因为蛋白denature得不够好。可以适当在loading buffer里增加sds浓度。
由于loading buffer与running buffer之间在表面活性上有差异，可以造成条带的扭曲
。为此，可以选用taurin buffer（代替glycin buffer）以弱化条带扭曲。甚至，有的
pi曾经建议，在上样之前，空跑至少10分钟。
用一般的样品跑胶，那么胶的品牌与型号之间的优劣不是很明显。你的sample比较特殊
，可能就放大了各厂家产品之间 |
|
s******y
发帖数: 28562 | 36 FT! FT! FT!
Of course you cannot ligate the single strand DNA like this way! Because
there is NOTHING to hold these two fragments together to allow the ligase to
act.
If you really want to ligate the single strand DNA, at least you should use
some kind of carrier system like BSA or other stuffs like a non-specific DNA
-binding protein.
If you choose to ligate from the double strand DNA, you can of course break
it down to single strand by incubating them on high temperature with
denaturing reagen |
|
T**********t
发帖数: 1604 | 37 哦，我没比较过，因为我一般不需要把结合上去的蛋白再洗下来。
我就是看说明书上这么说的：
2.2 Release of Immobilized Biotinylated Molecules
The biotin-streptavidin bond is broken by harsh conditions. 5 mins
incubation at 65°C or 2 mins at 90°C in 10 mM EDTA pH 8.2 with 95%
formamide will typically dissociate >96% of immobilized biotinylated DNA.
Alternatively, boil the sample for 5 mins in 0.1% SDS for
protein dissociation.
Please note that proteins will be denatured by such treatment and Dynabeads
Streptavidin can not be re-used.
It has al |
|
h*****d
发帖数: 210 | 38 加蛋白probe是不一样的， it is smear, but there is not a single distinct
probe. Could have some reasons for this, first of all, probe denaturing and
annealing is not good, I bought oligonucleotide, because it is highly
purified. the free probe is also smear which is about half length of the
smear with protein.
secondly, when I extract nuclear protein, I just use 250M sucrose in the
extraction buffer for nuclei and no further gradient centrifuge thereafter.
High/low salt method for getting nuclear protein( |
|
s******y
发帖数: 28562 | 39 It's normal that the protein order from the company may have precipitation
or denatured fractions during storage. Those will stuck in the resin.
Therefore, a 70% recovery rate is not bad.
P.S....."千老之王"...Me Ft... |
|
r*******i
发帖数: 112 | 40 glycerol 是蛋白稳定剂（因蛋白而异）并能保护蛋白在急冻时候少受破坏 >=5%才有用
imidazole 是弱的denaturant, 很多蛋白不喜欢
EDTA 是金属chelator, 如果你的蛋白里没有金属,应该没有伤害,但也没什么用,因为在
提纯中用EDTA 可以抑制金属蛋白酶
很多时候100mM KCl (or NaCl, (NH4)2SO4)with/without 50 mM Arg 增加可溶性和稳
定性, 原理很有争议, 也有用 Arg + Asp
最后, 浓缩,离心,分装,急冻 (N2 liquid), -80
or 50%glycerol -20 |
|
H*g
发帖数: 2333 | 41 I guess the author should have already included controls in gel as well as
western before posting this thread.
Could you please list some references for plant protein is harder to be
denatured, like under 4min@95C, or
3min@BoilingTemp?
gel
untranformed
or
that |
|
s******y
发帖数: 28562 | 42 Sorry I cannot give out any clear reference for plant protein being
difficult to denatured.
The thing is plant cells have cell walls, that's very different from
animal cells. Many proteins in plant cells are also N-oligo-glycosylated,
which is quite different from animal cell.
That idea was based on what people told me when I used to work in
a plant biology lab long time ago and I assumed that is a popular
assumption in the plant biology field. Maybe someone in this board
who is currently workin |
|
T**********t
发帖数: 1604 | 43 heat denature and snap cool may help
double- |
|
s******y
发帖数: 28562 | 44 denatured protein contaimination |
|
h*****e
发帖数: 1330 | 45 如果denature可以的话，加SDS后加热应该可以溶掉collagen，否则用collagenase了 |
|
V********n
发帖数: 305 | 46 250KD蛋白太大了，如果再来点高级结构很难弄的。这个试验很难搞啊。可以试试UV-
crosslink，然后跑denature的胶。 |
|
T**********t
发帖数: 1604 | 47 怎么recycle啊？streptavidin-biotin的binding很紧的，PCR都掉不下来，要洗下来除
非吧streptavidin denature了，可是那样一来不就没法recycle了？ |
|
m**i
发帖数: 47 | 48 我看了NEB家的elution的条件是75C的Tris-HCl （pH 7.5)，也许streptavidin
denature了等凉了之后还能在refold? pierce家的让用Guanidine-HCl,肯定就没法
recycle了。 |
|
s******y
发帖数: 28562 | 49 NO. Trypsin is a protease and won't bother with RNA.
Also, in most RNA extraction kit, there will be a step to denature all the
protein, that includes protease. |
|
