Acetyl Lysine Antibody, Agarose -2mg
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| Product Description | The acetylated lysine antibody is immobilized to agarose beads via amide linkages. The product can be utilized as an affinity matrix for rapid isolation and purification of proteins/peptides with acetyl lysine residues. | The acetylated peptides eluted from acetyl-lysine-specific affinity chromatography (ICP0388). Approximately 5 mg of trypsinated crude protein from TSA treated MMRU cells wereloaded. ![]() The maximum binding of acetylated BSA with ICP0388-2MG. -50 µl of ICP0388-2MG were incubated with 1 mg of acetylate BSA in a 1 ml tube for 60 min. - After washing with PBST 4 times, the bound acetylated BSA was eluted with 1 ml of 0.5 M HCl -5 µl / lane of the eluted acetylated BSA was resolved by SDS-PAGE and blotted wih monoclonal Mouse anti-Acetylated Lysine (ICP0390) | |
| Formulation | 0.5 mL of agarose beads suspended in 1 mL of slurry | ||
| Antibody Immobilized | 4 mg/ml antibody is covalently linked through amide bonds with NHS activated-SMCC, which further conjugates to thiolated agarose beads via thiol ether bonds. | ||
| Specificity | The antibody selectively captures proteins/peptides wtih acetylated lysine residues (N-epsilon). No cross reactions to methylated proteins/peptides. | ||
BindingCapacity | Approximately 0.2 mg of acetylated histone / mL slurry. | ||
| Applications | Rapid isolation and purification of proteins/peptides with acetylated lysine residues from cell lysate or protease-digested mixtures. | ||
| Scientific Description | Protein acetylation is a form of post-translational modification known to regulate many diverse biological processes. Detection, isolation and identification of acetylated proteins/peptides is essential in proteomic studies. Affinity chromatography is one of the most efficient and rapid methods to enrich and purify the acetylated species for further MS/MS identification. | ||
| Storage & Stability | Product is stable for 30 days at room temperature. For extended storage, store product at -20°C. Shake thoroughly before use.Expiration date is one year from date of shipping if properly stored. | ||
| Product Specific References |
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1. 血细胞与血清分离:
取人血液 1000ml,放置10min, 1000rpm离心20min .弃沉淀,留上清备用(沉淀为血细胞,上部为血清).
2. 乳糜粒分离:4000rpm 10°C离心10分钟,采用密度梯度离心
梯度液配置:离心管下部3/4容积加血浆,上部1/4容积加0.5MnaCl+0.3MEDTA,PH7.4 乳糜粒上浮,将乳糜粒吸出,留其余液体备用.
3. 血清蛋白分离:除去球蛋白,白蛋白及其它蛋白质.
①在用超滤除白蛋白,IgG中硫酸铵盐时,膜包该如何选择?选择几个?
②如用凝胶过滤来分级纯化血清中65%硫酸铵盐析出的白蛋白和纯化33%硫酸铵盐析出的IgG,其填充介质应选择什么?其洗脱缓冲液应用什么好?
下面有些关于这方面的数据,供参考:
①牛血清白蛋白:分子量:66210;分子形状:椭圆形;分子大小:
40Å*140Å;等电点:4.7;血浆中的含量:52.0g/L。
②IgG:分子量:15300;分子形状:球状;等电点:5.8—7.3;血浆中含量:2.0g/L。
③另外,我从书上看到说:凝胶过滤在分级方法中分辨率为中等,但对脱盐效果优良;流速较低,对分级每周期约≥8小时,对脱盐仅30分钟;适用于大规模纯化的最后步骤,在纯化过程的任何阶段均可进行脱盐处理,尤其适用于两种缓冲液交替时。
期待您的帮助,谢谢您。
1.以沉淀形式保存在高浓度的硫酸铵中。
2.添加50%甘油冷冻,尤其适用于酶类。
3.如果样品要用于生物学检测,避免使用防腐剂。在体内实验中不能添加防腐剂,而应当将样品分装成小份冷冻。
4.可使用无菌滤器以延长保存时间。
5.添加稳定剂,如甘油(5-20%)、血清白蛋白(10mg/ml)、配基(浓度取决于活性蛋白的浓度)以帮助维持生物活性。
6.避免反复冻融或冻干重溶解过程,这样很可能会降低生物活性。
7.一些冷沉淀蛋白会在4度沉淀出来,包括一些小鼠IgG3亚族的抗体,不能保存在4度冰箱中,可添加防腐剂保存在室温中。


