计算溶液所需的质量、体积或浓度。
BioReagent 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
2-8°C储存,禁止冷冻。低温运输,禁止冷冻 。请查阅批次 COA 获取详细规格。
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
储存缓冲液:20% (v/v) Ethanol
本产品采用重组Protein G蛋白作为配基,新型高刚性琼脂糖白球作为基质。Protein G可特异性地与抗体的Fc片段进行结合,本品可用于抗体(单抗和多抗)的纯化。经一步纯化即可从动物腹水、血清和培养液中获取高纯度的抗体。由于Fab区段有助于抗体结合,Protein G亲和层析介质更适合分离免疫复合物。Protein G在同时具有高载量、良好的理化稳定性、配基不易脱落,使用寿命长等优点。
Protein G相对Protein A对于IgG具有不同的结合特性,主要表现在Protein G更易与多克隆抗体进行结合。
阿拉丁rProtein G 亲和层析介质储存在20%乙醇中,凝胶和保护液的体积比为1:1,我司产品规格为实际凝胶的体积。
| 参数 | 指标 |
| 基质 | 高刚性琼脂糖 |
| 配基 | 重组Protein G |
| 平均粒径 | 75 μm |
| 配基密度 | 6-10mg Protein G/mL 介质 |
| 动态载量 | ≥35 mg h-IgG/mL 介质 |
| 最高流速(25℃) | 300 cm/h |
| 推荐流速 | <150 cm/h |
| 工作pH | 3-9 |
| 在位清洗pH | 2-10 |
使用说明
鉴于本品对不同种属、亚型的抗体具有不同程度的亲和性,保持不同抗体生物活性的所需缓冲条件不同,建议用户进行方法筛选。
以下提供一种参考工艺:
1、平衡:使用10 CV的buffer A(20 mM PB+0.15 M NaCl,pH 7.0)进行柱子平衡(平衡至基线);
2、上样:上样适量样品(固体样品请使用buffer A配制,液体样品可使用buffer A进行透析);
3、洗涤:使用5 CV的buffer A洗涤柱子(淋洗至基线);
4、洗脱:使用buffer B(20 mM柠檬酸,pH 3.0;或0.1 M甘氨酸,pH 3.0;或20 mM乙酸钠,pH 3.0)进行10 CV线性洗脱,直至100%B;
5、中和:收集洗脱产物,使用碱性缓冲液(1.0 M Tris-HCl,pH 9.0;或其它)将抗体稀释至稳定pH值;
6、再生:在进行一定次数的使用后,需要对柱子进行再生、原位清洗,以下提供两种方法:
(1)用0.1 M的乙酸或0.1 M的乙酸/20%乙醇淋洗柱子3-5 CV,然后用缓冲液洗到中性即可重复使用;
(2)也可用70%乙醇或6 M盐酸胍淋洗柱子3-5 CV,并用3-10 CV的纯水冲洗,然后用缓冲液洗到中性即可重复使用。
This product utilizes recombinant Protein G as the ligand and a novel high-rigidity agarose bead as the matrix. Protein G specifically binds to the Fc region of antibodies, making this resin suitable for the purification of both monoclonal and polyclonal antibodies. High-purity antibodies can be obtained in a single step from animal ascites, serum, or cell culture supernatants. Due to its additional binding capability via the Fab region, Protein G affinity resin is more suitable for isolating immune complexes. It offers high binding capacity, excellent physicochemical stability, minimal ligand leakage, and a long service life.
Compared to Protein A, Protein G exhibits distinct IgG binding characteristics, particularly a stronger affinity for polyclonal antibodies.
Aladdin rProtein G Agarose Resin is stored in 20% ethanol, with a settled gel to storage solution ratio of 1:1. The product specification refers to the actual volume of the settled gel.
| Parameter | Value |
| Matrix | High-rigidity agarose |
| Ligand | Recombinant Protein G |
| Average Particle Size | 75 μm |
| Ligand Density | 6–10 mg Protein G/mL resin |
| Dynamic Binding Capacity | ≥35 mg human IgG/mL resin |
| Maximum Flow Rate (25°C) | 300 cm/h |
| Recommended Flow Rate | <150 cm/h |
| Working pH Range | 3-9 |
| Cleaning-in-Place (CIP) pH Range | 2-10 |
Protocol
Due to the varying affinity of the resin for antibodies from different species and subtypes, and the differing buffer conditions required to maintain antibody activity, users are advised to optimize binding and elution conditions.
A reference purification procedure is provided below:
1.Equilibration: Equilibrate the column with 10 column volumes (CV) of Buffer A (20 mM phosphate buffer + 0.15 M NaCl, pH 7.0) until the baseline stabilizes.
2.Loading: Load the sample. For solid samples, dissolve in Buffer A. For liquid samples, dialyze against Buffer A prior to loading.
3.Washing: Wash the column with 5 CV of Buffer A until the UV baseline returns to baseline levels.
4.Elution: Elute bound antibodies using a linear gradient over 10 CV to 100% Buffer B (e.g., 20 mM citrate, pH 3.0; 0.1 M glycine, pH 3.0; or 20 mM sodium acetate, pH 3.0).
5.Neutralization: Collect elution fractions and immediately neutralize using an alkaline buffer (e.g., 1.0 M Tris-HCl, pH 9.0, or other suitable buffer) to adjust the antibody solution to a stable pH.
6.Regeneration & Cleaning-in-Place (CIP): After multiple cycles, regenerate the column using one of the following methods:
Option 1: Wash with 3–5 CV of 0.1 M acetic acid or 0.1 M acetic acid/20% ethanol. Rinse with buffer until neutral pH is reached before reuse.
Option 2: Wash with 3–5 CV of 70% ethanol or 6 M guanidine hydrochloride. Rinse with 3–10 CV of purified water. Equilibrate with buffer to neutral pH before reuse.
通过匹配包装上的批号来查找并下载产品的 COA,每批产品都进行了严格的验证,您可放心使用!
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | P749485 | |
| 分析证书 | P749485 |