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。
本产品由新型高刚性琼脂糖基质和rProtein A配基组成。rProtein A 通过大肠杆菌表达、纯化得到,生产过程无动物来源物料风险。新设计的rProtein A相比天然Protein A 拥有良好耐碱性、与IgG Fc片段更强的特异性结合能力的特点。新型高刚性琼脂糖基质具有良好的化学稳定性和生物相容性。采用定点偶联技术将rProtein A固定在高刚性琼脂糖基质上,使之具有载量高、流速高、理化稳定性好、非特异性吸附低、配基脱落少,使用寿命长等优点。本品可以满足从腹水、血清和细胞培养液等样品中一步纯化得到高纯度抗体的工艺需求,是大规模生产抗体的理想选择。
阿拉丁耐碱rProtein A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 | 耐碱重组Protein A |
| 平均粒径 | 70 μm |
| 动态载量(6min保留时间) | 65~80 mg h-IgG/mL 介质 |
| 最高流速(25 ℃) | 300 cm/h |
| 配基脱落 | <10 ng/mL |
| 最高耐压 | 0.3 MPa (3 bar) |
| 在位清洗 | 0.1~0.5 M NaOH溶液 pH 2~14 |
| 保存条件 | 2~8℃(20%乙醇)或(2%苯甲醇) |
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5-10 CV的buffer A(20 mM PB+0.15 M NaCl, pH 7.0;或 0.05 M 硼酸,4.0 M NaCl, pH 9.0)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。
4、中和:收集洗脱产物,使用中和缓冲液(1.0 M Tris-HCl, pH 9.0;或其它)将抗体稀释至稳定pH值。
5、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0.1-0.5 M NaOH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 is composed of a novel high-rigidity agarose matrix coupled with recombinant Protein A (rProtein A) ligand. The rProtein A is expressed and purified from E. coli, ensuring an animal-origin-free production process. The newly designed rProtein A exhibits improved alkali resistance and stronger specific binding to the Fc region of IgG compared to native Protein A. The high-rigidity agarose matrix offers excellent chemical stability and biocompatibility. Through site-specific coupling technology, rProtein A is immobilized onto the matrix, resulting in high binding capacity, high flow rate, superior physicochemical stability, low non-specific adsorption, minimal ligand leakage, and extended service life.
This resin is suitable for one-step purification of high-purity antibodies from samples such as ascites, serum, and cell culture supernatant, making it an ideal choice for large-scale antibody production.
Aladdin Alkali-tolerant rProtein A 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 | Alkali-resistant rProtein A |
| Average Particle Size | 70 μm |
| Dynamic Binding Capacity (6 min residence time) | 65–80 mg human IgG/mL resin |
| Maximum Flow Rate (25°C) | 300 cm/h |
| Ligand Leakage | <10 ng/mL |
| Maximum Pressure Tolerance | 0.3 MPa (3 bar) |
| Cleaning-in-Place (CIP) | 0.1~0.5 M NaOH pH 2~14 |
| Storage Conditions | 2–8°C in 20% ethanol or 2% benzyl alcohol |
Protocol
Due to varying affinity of the resin for antibodies from different species and subtypes, and differences in buffer conditions required to maintain antibody bioactivity, method screening is recommended.
A reference purification procedure is provided below:
1.Equilibration: Equilibrate the column with 5–10 column volumes (CV) of Buffer A (20 mM phosphate buffer + 0.15 M NaCl, pH 7.0; or 0.05 M borate, 4.0 M NaCl, pH 9.0) 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 (20 mM citrate, pH 3.0; or 0.1 M glycine, pH 3.0; or 20 mM acetate, pH 3.0).
5.Neutralization: Collect the elution fractions and immediately neutralize using a neutralization buffer (e.g., 1.0 M Tris-HCl, pH 9.0, or other suitable buffer) to bring 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 0.1–0.5 M NaOH. 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 | P405880 | |
| 分析证书 | P405880 |