Dojindo,IgG纯化试剂盒-G/1/AP02,IgG Purification Kit-G

IgG 纯化试剂盒用于山羊、小鼠、兔和其他动物的免疫球蛋白 G 的分离和纯化。 该试剂盒包含固定的蛋白 A 或 G 和缓冲溶液,可最大限度地回收 IgG。 从血清或其他含有 IgG 的溶液中分离和纯化 IgG 的总时间约为 30 分钟(图 1)。 由于蛋白 A 或 G 的支持物是硅胶,离心后凝胶上保留溶液的体积非常小。 因此,所有不与蛋白A结合的蛋白质或其他物质都可以通过洗涤两次来去除。 图 2 显示了来自各种动物的纯化 IgG 的 SDS-PAGE。此外,在洗脱过程中 IgG 的变性是最小的,因为与凝胶结合的 IgG 会以非常快的过程释放。 本试剂盒中的凝胶可重复使用 20 次以上,性能相同。 用过的凝胶在 0-5ºC 的洗涤缓冲液中也可稳定保存一年。

Fig. 1 IgG isolation process

Fig. 2 SDS-PAGE of isolated immunoglobulin by using IgG PurificationPurification Kit-A and Kit-G.WS : Whole SerumA : Prepared by IgG Purification Kit-AG : Prepared by IgG Purification Kit-GCondition: 6% acrylamide gel/Tris-glucine buffer

PrecautionIf the IgG solution contains gelatin,before applying the solution to the kit, enzyme digestion may be required.

Table 1 IgG Recovery from 50 μl Serum

What is the recovery of IgG with this kit?

It depends on the type of IgG and type of animal. In the case of high affinity type IgG for protein A or protein G, about 70-80% IgG is recovered from 100-200 μg IgG or IgG solution containing other proteins or macromolecules.

How is the purity of purified IgG using this kit?

The purity of the IgG from various serum is indicated in the figure above. Highly purified IgG is available with only a one time purification process.

How much IgG can be recovered from serum?

About 150-350 μg IgG can be recovered from 50 μl serum. I-7. Protein Labeling: IgG purification

How many times can a protein A gel be used?

At least 20 times.

Can I use the used protein A gel or protein G gel for the purification of different IgG solutions?

Use a new gel for a different sample to avoid contamination.

Is a used protein A gel or protein G gel stable?

A used protein A gel or protein G gel in Washing buffer is stable at 0-5ºC for one year.