a****t
发帖数: 2 | 1 Please take a look at http://www.ab-mart.com
Abmart is the largest custom antibody developer with the capacity to develop
2000 mAb/year. It provided superior quality with great price. |
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b******c
发帖数: 38 | 2 Thank you so much!
I've searched on Bio-Rad's, and the product "Rabbit Anti-Goat IgG (H+L)-HRP
Conjugate #172-1034" popped out. However, it's said to be an ELISA antibody.
Did you guys also use it for Western blotting? I really appreciate your
help |
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h*******o
发帖数: 4884 | 3 Re-use
Use Casein or Pierce Superblock to dilute antibody, so it can be re-used and
stored at 4oC for longer time without precipitation
Also, use signal enhancer |
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a*****g
发帖数: 543 | 4 before this, please use enough antibody to cover the whole blot to make sur
e there is no other interesting/relevant bands...
1 |
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l*****m
发帖数: 122 | 5 最近需要在老鼠里杀掉DC看看 但是又不能用DTR来做 想看看有没有直接用antibody
deplete DC的应用 我查到的基本都是in vitro条件的 想问问大家有没有谁做过in
vivo的呀？或者有推荐文献也行 谢谢啦！ |
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s****9
发帖数: 932 | 6 It is unlikely to use antibody to deplete DC in vivo. |
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w******e
发帖数: 1187 | 7 比如：有一target有highly homologous protein（且称negative target），如何
改造antibody以确保specificity（不bind negative target）？ |
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w******e
发帖数: 1187 | 8 to answer this question, let's do a "simple" math 1st:
Say you generated polyclone antibody which let's say has only two Abs -- X (
the
"nonspecific" one), Y (the "specific" one). the ratio b/w X and Y is 100:1.
the affinity of both abs against the real target P is 1nM, while the
affinity
against the homolog Q is 10nM for X, but 1uM for Y (hence the specificity
difference). You want to add excessive amount of Q to soak off Ab X so that
the final ratio b/w X and Y is 1:10 instead of 100:1, and th... 阅读全帖 |
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H****s
发帖数: 301 | 9 现在这些已经快过时了。多读读antibody engineering最近的综述吧。 |
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H****s
发帖数: 301 | 10 很抱歉。这两天比较忙，关于抗体工程这个领域，一时半会说不清楚，因为不仅仅是学
术研究问题。如果你对这个领域有点兴趣，建议你pubmed一下以下几个人写的综述：
Richard Lerner，John McCaffery，Geoege Smith， Gregory Winter， Andrew
Bradbury和James Marks。这几个人基本上是antibody phage/yeast display领域的开
创者，并且也是抗体工程以后发展的方向。如果你对某一个人特别感兴趣，请告诉我，
我再详细给你说关于这个人的一些情况（学术研究方面的）。 |
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s****9
发帖数: 932 | 11 No worry. Antibody is very stable. Without bacteria contamination, it can
be stable in room temperature for half a year. |
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m*p
发帖数: 226 | 12 Different antibodies are quite different. Whether it works or not, it can be
easily tested by a WB or IHC. |
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S****r
发帖数: 982 | 13 Anybody has experience and glad to share which antibodies of CBP/CREBBP,
p300, and GRIP1 are good for western blotting and CHIP. Many thanks! |
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m******5
发帖数: 1383 | 14 I found some claimed to be 'Chip qualified' antibody naturally gave out
super strong background (when using non transfected or knock out cell line
as negative control), maybe due to non specific binding to DNA or Histon?
I start to wonder if those paper simply use IgG as negative control
really got validated result. |
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d****7
发帖数: 109 | 15 嗯，确实，比如Abcam的GFP antibody (ab290)，跑western看一眼，好多好多非特异带
，结果N多人用这个做ChIP-seq |
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s********n
发帖数: 248 | 16 ECL,take the ECL out when you are done with 2nd antibody, prewarm in room temp for about 25min, then use 4ml(buffer A)100ul(buffer B) for a 6*4cm^2 membrane, 4min, then develop. |
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l*****a
发帖数: 1431 | 17 现在用sigma的mouse monoclonal anti-flag antibody （M2），总是有很多非特异性
的bands 而且和我的band挨得很近。请有经验的xdjm给推荐个干净点的anti-flag。多
谢！ |
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l*****a
发帖数: 1431 | 18 谢谢LS。是 stratagene anti-flag M2 antibody, Cat#200472 吗？ |
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f******9
发帖数: 39 | 19 Thanks in advance.
Please recommend the good antibodies for 3 protein: Alix, TSG101, CD9. The
main purpose is to do western blot. |
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m******5
发帖数: 1383 | 20 someone tell me the F-7 probe is the best amoung all the available whole
mount immune staining antibody, I am trying that, but I also need to buy
some other strain for back up |
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m******5
发帖数: 1383 | 21 could you send me the CATog number of the secondary antibody you used?
BTW, what is your original purpose of doing whole mount immunestaining? |
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n***w
发帖数: 2405 | 22 I used mouse monoclonal HA antibody from Millipore which is not bad since I
had no experiences with other suppliers. |
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l**********9
发帖数: 264 | 23 Does anyone know if there is a good anti-Venus antibody used on pariffin-
embedded sections (IHC)?
Thanks |
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A******y
发帖数: 2041 | 24 Most antibody is generated using peptide, in your case, it is probably
generated from phos-peptide. Are you detecting protein or just peptide? If
just peptide, using mass-spec is probably easier. |
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c**********e
发帖数: 230 | 25 以往大家问了好些次, 都是做WB, 我想问问做immunostaining的flag antibody, 不知
道大家有没有推荐?
多谢多谢 |
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a****d
发帖数: 1919 | 26 M2 Flag antibody from Sigma works well for immunostaining. |
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C*******e
发帖数: 4348 | 27 可以包子求一个用flag antibody做IP的protocol么
过段时间可能会用到 |
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g*********5
发帖数: 2533 | 28 I have peptide sequence and need rabbit polyclonal antibody sevice
whcih company is good for price/quality?
my tried biosyn, covance. feel expensive, and quality so so.
thanks. |
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p******d
发帖数: 3737 | 29 不是GFP，是YFP的antibody，做IF。实验室里面有的都是主要做GFP的，试了一下，不
太好。 |
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s**********y
发帖数: 80 | 31 大家elisa用的antibody一般都是从什么公司买的啊？最近需要买一点，哪位知道好的
推荐推荐， 谢谢 |
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d**l
发帖数: 1546 | 32 We buy ELISA kits from R&D Sys, Promega, CST, etc.
It takes time to buy antibodies and optimize an ELISA |
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F******p
发帖数: 2099 | 33 I used Cell Signaling antibody for NF-kB and phos-NF-kB, very weak signal
even in stimulated RAW cells. Can somebody please recommend good NF-kB
antiboies, especially phos-NF-kB and Acylated NF-kB.
Thanks a lot! |
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h*****G
发帖数: 113 | 34 大家好。
我需要做triple fluorescence immunostaining.
在红色和绿色channel分别用了Alexa 488和Alexa 594, 求推荐能用在Blue channel的
2nd antibody.
我们实验室显微镜是zeiss 的observer D1, blue channel的filter set 是zeiss的
filter set 49.
谢谢。 |
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h*******o
发帖数: 4884 | 35 catalase antibody 好像很容易找吧
连abcam这种公司做出来的都还行,我以前用过
楼主不如找个常用的公司买一个就行了
而且个人经验,花2倍,3倍的钱买到一个好抗体都比买个便宜的不太work的划算好多 |
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P****o
发帖数: 172 | 36 just bought one antibody from this supplier, it works. |
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g*********5
发帖数: 2533 | 37 thanks.
what the antibody name?
I want buy alas2.
the santa cruz and sigma one not work...
and Novus one's work in a paper, but its peptide include mitochondria
leading peptides... |
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y*****w
发帖数: 146 | 38 现在在做yeast surface display， 大家能帮忙推荐一个好的Anti-HA PE conjugated
Monoclonal Antibody 吗？试了miltenyibiotec 的Anti-HA-PE， 信号很不好。 十分
感谢！ |
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F*******1
发帖数: 124 | 39 Is there any macrophage marker that is not found in Kupffer cells? I am
looking for an appropriate antibody to target macrophage surface marker
without recognizing the macrophages in the liver. Thanks! |
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h*******o
发帖数: 4884 | 40 Same here with MBP antibodies from a couple of different vendors
I think it is quite normal for MBP to have multiple bands in WB. |
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n***w
发帖数: 2405 | 41 看paper上用的基本都是R&D的mAb，可是我想染老鼠组织嗯，所以想避开monoclonal
antibody.
大家有没有什么经验，还有哪些公司的Ephrin-B4好使用于组织染色。
谢谢。 |
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i***0
发帖数: 160 | 42 I have tried increase wash volume, wash time, times of wash, adjusting
antibody concentration. Occasionally, I got a relatively clean image, but
still not clean. Someone told me I could try add some salt in PBST for wash,
and I am trying it today. I will report my result when I got it. But if you
have any great tricks, please help. Thanks |
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i***0
发帖数: 160 | 43 Thank you very much! Currently I am using 5% milk and 10^4 times of dilution
for secondary antibody.But I will try BSA next time. I have tried primary 1
hr at RT and O/N, and it seems better with primary 1 hr. I just hate
trouble shooting. |
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a*****g
发帖数: 543 | 44 Thermo V5 antibody is as goog as Invitrogen (life tech...) for western blot.
not sure other applications |
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y******8
发帖数: 1764 | 45 Invitrogen's V5 antibody is Monoclonal.
V5, |
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y****n
发帖数: 332 | 46 Hi, i want to test if a protein is methylated. is there a good antibody to
test it? Thanks! |
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c*********7
发帖数: 148 | 47 有木有用试了才知道？不过你一上来就要用antibody是不是工程略显浩大了点。。。 |
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C*******e
发帖数: 4348 | 48 不是啊，我就是问问大家这个方法啊
这个方法本来就是一上来就是antibody啊
还是驼羊的
然后又是建库，又是phage display筛选，再共表达
工程就是很浩大啊
所以实在没有办法“有没有用都试一下先”。。。。。 |
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s**********y
发帖数: 80 | 49 Anyone can suggest good human MMP-2 antibody and recom protein? Thanks! |
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n***w
发帖数: 2405 | 50 hello，有哪位大侠知道好用的agrin antibody, 最好是wb和ihc（mouse）都可以的。
。。
另外，endothelial markers无外乎cd31, cd34, endomucin, meca32, etc....但是在
我的ko里，cd31貌似是change的，所以我想找一个不太可能变化的marker，求大侠们推
荐以及探讨。。
谢谢。 |
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