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1楼主 = =2012/7/9 18:22:00
自己在DXY那边也求助问了，没人理我
被亲友提醒，于是试试求助潜伏的叉球的各路大神
俺是自己合成多肽，间接法ELISA测血清抗体
之前的板子不好用，要换新板子
目前锁定了NUNC的96孔
现在问题来了，我的多肽亲水性特别强（负的2点多）
NUNC的表面有maxisorp和multisorp两种可用
但是！！！前面的差点，有现货，后面的好点没现货
从国外调货的周期是两个月啊两个月
买有现货的吧，要买就必须买一整箱！！！60块啊，试了下不好用的话我拿回家数米粒减肥用吗？！不敢买
走投无路了这边问问碰碰运气
有没有GN做过类似的实验，maxisorp能包上不？
BT没打完，花式是瘦十斤好offer好男人！！！
先谢过
9 = =2012/7/10 3:31:00
看了一下Thermo的官網
- PolySorp?, with high affinity to molecules of a hydrophobic nature
- MediSorp?, has a surface chemistry between PolySorp? and MaxiSorp?.? Provides low background signal with samples containing serum.
- MaxiSorp?, with high affinity to molecules with mixed hydrophilic/hydrophobic domains
- MultiSorp?, with high affinity to hydrophilic molecules
http://www.nuncbrand.com/en/page.aspx?ID=241
What is the difference between PolySorp, MaxiSorp, and MiniSorp surfaces?
MaxiSorp, PolySorp and MiniSorp surfaces were developed for immunology assays. The MaxiSorp surface is a modified, highly charged polystyrene surface with high affinity to molecules with polar or hydrophilic groups. The surface has a high binding capacity for proteins, including globular antibodies in proper orientation. Thus, it offers very high sensitivity in double antibody "sandwich" tests. The PolySorp surface is more hydrophobic than the MaxiSorp surface. It has high affinity to molecules of a more hydrophobic character. This surface is particularly suited to non-protein antigens including virus antigens. The MiniSorp surface is a polyethylene surface with very low affinity to molecules of any type. This type of surface is ideal for the liquid phase immuno techniques. The MiniSorp surface is only available in a tube format, while the MaxiSorp and PolySorp surfaces are offered in both 96-well plate and module formats.
http://www.nuncbrand.com/en/page.aspx?ID=298
個人覺得看官網說明LZ選maxisorp好像也還可以,而且實驗室有其他人做疏水性蛋白也能用,
另外如果選multusorp, LZ確定blocking reagent (如BSA)能順利binding嗎?
另外如果是background太高,可能能從modify protocol下手,
如降低抗原濃度,降低抗體濃度,增加blocking reagent濃度和blocking時間,增加wash buffer的detergent濃度或wash次數等...
祝好運!!
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