虾血蓝蛋白 (SHC) 是一种源于对虾的高可溶性血蓝蛋白，可替代钥孔帽贝血蓝蛋白 (KLH)，作为载体蛋白与多肽、核酸、药物或毒素等低分子量分子偶联，使其具备高免疫原性。经测试，SHC 具有与 KLH 相当的免疫原性，而其溶解度 ( 尤其是在与半抗原偶联后 ) 则显著高于 KLH，从而使免疫原制备方案具有更大的灵活性。产品形式：血蓝蛋白以 PBS 溶解的冻干形式提供。使用时，可加入超纯水重悬。
产品特点
高免疫原性：可完美替代 KLH；
高得率偶联：偶联效率与 KLH 相同；
高溶解度：溶解度显著高于 KLH，兼容不常见或溶解性很差的半抗原；
高纯度：纯度大于 98%；
性能卓越：已经过数百例抗体制备验证；
高性价比：价格远低于 KLH。
使用方法
SHC 活化及半抗原偶联
SMCC 活化
1. 使用适量超纯水将 SHC ( 未活化 ) 冻干粉复溶为 20 mg/mL 的 SHC 溶液；
2. 向 1 mL 复溶后的 SHC 溶液中加入 800 µL 超纯水配制的 SMCC 溶液 ( 5 mg/mL，现配现用 )；
3. 置于混匀仪上温和翻转，室温孵育 60 min 或 37℃孵育 30 min；
4. 使用分子筛层析除去过量的 SMCC。
与半抗原偶联
1. 将 20 mg 含有巯基的半抗原溶解于 5 mL 偶联缓冲液 (83 mM sodium phosphate, 0.1 M EDTA, 0.9 M NaCl, 0.1 mM TCEP, pH 7.2) 中；
注意：0.1 mM TCEP 不是必需的，它的主要作用是将半抗原的二硫键还原成巯基。
2. 立即将半抗原溶液与活化后的 SHC 按照质量比 1 : 0.6 混合，室温反应 2 h 或 4℃过夜反应；
注意：该偶联比例为推荐比例。不同半抗原大小和结构的差异会影响偶联效率，为确保更有效的偶联， 需根据具体情况，对半抗原和 SHC 偶联比例进行多次预实验验证。
3. 使用分子筛层析除去 EDTA。
EDC 活化与半抗原偶联
EDC 活化
1. EDC 和 NHS 在使用前需平衡至室温；
2. 使用适量激活缓冲液 ( 0.1 M MES, 0.5 M NaCl, pH 6.0) 将 SHC( 未活化 ) 冻干粉复溶为 1 mg/mL 的 SHC 溶液；
3. 向 1 mL 上步配制的 SHC 溶液中加入 0.4 mg EDC 和 0.6 mg NHS，混匀，室温反应 15 min；
4. 向反应液中加入 1.4 µL β- 巯基乙醇使 EDC 失活； 5. 使用分子筛层析除去过量的 EDC、NHS 和β- 巯基乙醇。
与半抗原偶联
1. 向活化后的 SHC 溶液中加入等摩尔量的半抗原 ( 溶解于 PBS 中 )，室温反应 2 h；
2. 向上步反应液中加入终浓度为 10 mM 的羟胺 (hydroxylamine) 终止反应；
3. 使用分子筛层析除去过量的羟胺。
注意事项
1. 本产品定量精确，请在原装管内按照所需浓度直接溶解使用；请勿对本产品进行分装，否则会因产品 性状 / 静电等原因造成较大损失；
2. SHC 预测消光系数ε1% 为 10，如需要测定 SHC 溶液浓度请调整消光系数；
3. 为了您的安全和健康，请穿实验服并戴一次性手套操作；
4. 本产品仅限科研使用。
Shrimp Hemocyanin (SHC) is a highly soluble hemocyanin derived from shrimp. It can be used as an alternative to Keyhole Limpet Hemocyanin (KLH) as a carrier protein for conjugation with low-molecular-weight molecules such as peptides, nucleic acids, drugs or toxins to render them highly immunogenic. Tests have shown that SHC exhibits immunogenicity comparable to KLH, while its solubility (especially after conjugation with haptens) is significantly higher than that of KLH, allowing greater flexibility in immunogen preparation protocols.Product Format: Lyophilized powder dissolved in PBS. Reconstitute with ultrapure water before use.
Product Features
High Immunogenicity: Perfect replacement for KLH;
High Conjugation Yield: Conjugation efficiency identical to KLH;
High Solubility: Significantly higher solubility than KLH, compatible with uncommon or poorly soluble haptens;
High Purity: Purity > 98%;
Excellent Performance: Validated in hundreds of antibody preparation cases;
Cost-Effective: Price much lower than KLH.
Usage
SHC Activation and Hapten Conjugation
SMCC Activation
1. Reconstitute lyophilized SHC (unactivated) with an appropriate volume of ultrapure water to a final concentration of 20 mg/mL;
2. Add 800 µL of freshly prepared SMCC solution (5 mg/mL in ultrapure water) to 1 mL of reconstituted SHC solution;
3. Incubate with gentle mixing for 60 min at room temperature or 30 min at 37 °C;
4. Remove excess SMCC by size-exclusion chromatography.
Conjugation with Hapten
1. Dissolve 20 mg of thiol-containing hapten in 5 mL of conjugation buffer (83 mM sodium phosphate, 0.1 M EDTA, 0.9 M NaCl, 0.1 mM TCEP, pH 7.2);
Note: 0.1 mM TCEP is optional; its main function is to reduce disulfide bonds in the hapten to thiol groups.
2. Immediately mix the hapten solution with activated SHC at a mass ratio of 1:0.6, and react for 2 h at room temperature or overnight at 4 °C;
Note: This conjugation ratio is recommended. Variations in hapten size and structure may affect conjugation efficiency. To ensure efficient conjugation, optimize the hapten-to-SHC ratio via multiple preliminary experiments.
3. Remove EDTA by size-exclusion chromatography.
EDC Activation and Hapten Conjugation
EDC Activation
1. Equilibrate EDC and NHS to room temperature before use;
2. Reconstitute lyophilized SHC (unactivated) with an appropriate volume of activation buffer (0.1 M MES, 0.5 M NaCl, pH 6.0) to a final concentration of 1 mg/mL;
3. Add 0.4 mg EDC and 0.6 mg NHS to 1 mL of the above SHC solution, mix well, and react for 15 min at room temperature;
4. Add 1.4 µL β-mercaptoethanol to quench EDC;
5. Remove excess EDC, NHS and β-mercaptoethanol by size-exclusion chromatography.
Conjugation with Hapten
1. Add an equimolar amount of hapten (dissolved in PBS) to the activated SHC solution, and react for 2 h at room temperature;
2. Add hydroxylamine to a final concentration of 10 mM to stop the reaction;
3. Remove excess hydroxylamine by size-exclusion chromatography.
Notes
1. This product is accurately quantified. Dissolve directly in the original vial at the desired concentration. DO NOT aliquot, as significant loss may occur due to product properties / static electricity;
2. The predicted extinction coefficient ε1% of SHC is 10. Adjust the coefficient when determining the concentration of SHC solution;
3. For your safety and health, wear a lab coat and disposable gloves during operation;
4. For research use only.
ZJ26F0433795
危险品化学品经营许可证（带存储）