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BioReagent,天然 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
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在色谱分析、有机合成和交叉偶联反应领域已被 4 篇同行评审文献引用。
本产品为海水养殖 KLH ,是一种纯化的钥孔帽贝血蓝载体蛋白,能够轻松地制备含有肽抗原的高效免疫原。
特性:
• 高得率偶联—CCH 的每个分子包含数百个通过 EDC 或 NHS 酯交联剂进行半抗原偶联的伯胺;
• 质量经验证—与传统的载体蛋白源不同的是,纯化和稳定的海水养殖 KLH 维持了在水溶液中的溶解度;
• 可持续来源—KLH 是从在海水养殖中生长的软体动物大锁孔帽贝(钥孔帽贝)的精选种群中分离得到的,而不是从野生种群中提取的;
• 高免疫原性—KLH 具有高分子质量(4.5×10⁵~1.3×10⁷ Da;350和390 kDa 亚基聚集体),可引发比 BSA 或卵清蛋白更强的免疫应答。
KLH 是较常用的具有免疫原性的载体蛋白,用于制备免疫和抗体生产所需的肽抗原。在磷酸盐缓冲液中的 KLH 制备可随时用于通过胺反应性 NHS-酯或戊二醛交联反应实现的半抗原-载体偶联。
载体蛋白是大而复杂的分子,能够在进样后刺激免疫应答。成功的生产特异针对小抗原的抗体(例如:肽段或药物化合物)要求这些半抗原被共价交联于更大、更复杂的分子(通常为某种蛋白质)上,以使其具有免疫原性。基于免疫原性、溶解度以及是否能够达到足够的载体偶联,选择载体蛋白。
出于抗体生产的目的,钥孔帽贝血蓝蛋白 (KLH) 广泛用作与半抗原和其他抗原偶联的载体蛋白,使其更具免疫原性。由于其质量大、复杂性高,与其他载体蛋白相比,KLH 能够引发更强的免疫应答。由于该蛋白分离自软体动物,因此与哺乳动物物种在系统发育上相距较远,并且不太可能产生会与测定中的典型靶点样品发生交叉反应的抗体。
本产品是从海水养殖而非野生生长的帽贝中分离得到的。具有较高的免疫原性;它在缓冲液中纯化和冻干以优化其稳定性和溶解性,可进行半抗原偶联。KLH 是一种蛋白大分子,拥有数百个伯胺和羧基基团,可以成为与 N-羟基琥珀酰亚胺酯、EDC 以及其他交联试剂偶联的靶点。
碳二亚胺交联剂 EDC 可将含羧基的半抗原(例如,肽抗原的 C 末端)偶联到高度免疫原性 KLH 载体蛋白上,用于免疫接种和抗体生产。这种免疫原制备的方法最适用于主序列中央区域含有较少或不含天门冬氨酸残基、谷氨酸残基(羧酸盐)和赖氨酸残基(伯胺)的肽抗原。由于肽含有羧酸盐和胺,EDC 偶联可使其形成各种聚合物,并在交联的载体蛋白上呈随机取向。通常情况下,这样可令载体蛋白上呈现高水平的抗原装载量,并且各个方向上均可诱导抗体产生。不过,抗原肽序列内的重要(所需的)抗原表位区域如果含有伯胺(赖氨酸残基)或羧酸盐(天冬氨酸和谷氨酸残基),该表位可能被 EDC 介导的偶联所阻断。在这种情况下,可以使用同源双官能团胺-反应性交联剂与磷酸盐缓冲液中的纯化KLH,或用独特的末端半胱氨酸合成肽以及马来酰亚胺活化 KLH 试剂盒来制备载体蛋白偶联物。
This product is Mariculture KLH, a purified keyhole limpet hemocyanin carrier protein that enables simple preparation of highly effective immunogens with peptide antigens.
Features:
• High-yield conjugation—each molecule of KLH contains hundreds of primary amines available for coupling haptens via EDC or NHS ester crosslinkers
• Validated quality—purified and stabilized mariculture KLH maintains solubility in aqueous solutions, unlike traditional sources of the carrier protein
• Sustainable source—KLH is harvested from select populations of the mollusk Megathura crenulata (keyhole limpet) that are grown in mariculture, not extracted from wild populations
• Highly immunogenic—KLH has a high molecular mass (4.5×10⁵~1.3×10⁷ Da; aggregates of 350 and 390 kDa subunits) and elicits a stronger immune response than BSA or ovalbumin
KLH is the most popular and immunogenic carrier protein used for preparation of peptide antigens for immunization and antibody production. KLH preparations in phosphate buffer are ready for hapten-carrier conjugation via amine-reactive NHS-ester or glutaraldehyde crosslinking.
Carrier proteins are large, complex molecules capable of stimulating an immune response upon injection. Successful production of antibodies specific to small antigens (i.e., peptides or drug compounds) requires that these haptens be covalently conjugated to a larger, more complex molecule (usually a protein) to make them immunogenic. Carrier proteins are chosen based on immunogenicity, solubility, and whether adequate conjugation with the carrier can be achieved.
Keyhole limpet hemocyanin (KLH) is widely used as a carrier protein for conjugation to haptens and other antigens to make them more immunogenic for the purpose of antibody production. Because of its large mass and complexity, KLH elicits a stronger immune response than other carrier proteins. Because the protein is derived from a mollusk, it is phylogenetically distant from mammalian species and less likely to produce antibodies that will cross-react with typical target samples in assays.
This product is KLH that has been harvested from limpets grown in mariculture rather than captured from the wild. It provides the highest possible immunogenicity; it is purified and lyophilized in buffers that optimize its stability and solubility for hapten conjugation. As a large protein, KLH has hundreds of primary amines and carboxyl groups that can be targeted for conjugation with NHS-ester, EDC, and other crosslinking reagents.
The carbodiimide crosslinker EDC conjugates carboxyl-containing haptens (e.g., C-terminus of peptide antigens) to the highly immunogenic KLH carrier protein for immunization and antibody production. This method of immunogen preparation is ideal for peptide antigens with few or no aspartic and glutamic acid residues (carboxylates) and lysine residues (primary amines) within the central portion of the primary sequence. Because peptides contain both carboxylate and amines, EDC conjugation results in their becoming variously polymerized and randomly oriented in their linkage to the carrier protein. Typically, this results in a high level of antigen loading on the carrier protein as well as presentation in all possible orientations for antibody production. However, important (desired) epitopes within the antigen peptide sequence may be blocked by EDC-mediated conjugation if those regions contain primary amines (lysine residues) or carboxylates (aspartic and glutamic acid residues). In these cases, either use a homobifunctional amine-reactive crosslinker with the purified KLH in phosphate buffer or synthesize the peptide with a unique terminal cysteine and use a kit with Malemide-Activated KLH to prepare the carrier protein conjugate.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | H1443352 | |
| 分析证书 | H1443352 | |
| 分析证书 | H1443352 |