CleanAb清除试剂盒
快速和简便地从偶联抗体中去除多余的寡核苷酸或小分子染料
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适用于Rabbit IgG或Mouse IgG（所有亚型）
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高效去除多余的Oligo或小分子染料
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温和条件洗脱（pH 7.5）
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回收率高达60%–90%
CleanAb清除试剂盒
CleanAb试剂盒可在温和条件下，快速、简便地去除Oligo偶联一抗体系中多余的寡核苷酸，适用于两种纯化方式：抗体偶联后纯化或抗体先结合beads再偶联纯化。
该试剂盒采用经过特殊处理的琼脂糖珠，可在pH 7.5的条件下实现温和洗脱，在高效去除杂质的同时，实现目标样品的高回收率。
CleanAb试剂盒适用于Rabbit IgG或Mouse IgG（包括所有亚型），可满足常规抗体类型的纯化需求。
试剂盒组分
- CleanAb Beads for Rabbit or Mouse IgG
- CleanAb Elution Buffer
- CleanAb Wash Buffer
- Spin columns
Immunostaining of HeLa cells with mouse anti-B23 antibodies (60096-1-Ig, 1:100, orange), on-bead chemically conjugated to a 15-nt ssDNA oligo, purified using CleanAB beads and desalted. Detection was performed by hybridization with an imaging DNA strand labeled with ATTO643.
CleanAb试剂盒产品列表
| 类型 | 识别Rabbit IgG货号 | 识别Mouse IgG货号 |
|---|---|---|
| Agarose | CK001 | CK003 |
| Magnetic Agarose | CK002 | CK004 |
适用范围
寡核苷酸去除
CleanAb试剂盒可高效去除寡核苷酸偶联抗体中的游离寡核苷酸，可适配15-90 nt多种常用长度及不同修饰类型的寡核苷酸，包含带有聚腺苷酸尾（Poly (A)-tail）、硫代磷酸修饰的寡核苷酸；可满足CODEX、PLA/PEA、NULISA 以及CITE-Seq等多种下游终端应用的纯化需求。
染料去除
CleanAb试剂盒能够有效清除染料偶联抗体的过量荧光染料，适用染料种类包括呫吨类染料（FITC）、花菁类染料（Cy3、Cy5）、Alexa Fluor™系列染料（Alexa Fluor™ 647、750）、Atto系列染料（Atto 647N）以及Aquora™系列染料（Aquora™ 488、550、650）。经本试剂盒纯化后的抗体偶联物已通过免疫荧光（IF）成像验证。注意：本试剂盒不适用于藻红蛋白（PE）或别藻蓝蛋白（APC）的去除纯化。
CleanAb操作流程-一个盒子，两种纯化方式可选
采用ATTO643标记的成像DNA链进行杂交，实现对CD137的IF检测
Mouse IgG (anti-Lamin B1 IgG1; 66095-1-Ig) was conjugated with NHS-Azide (lane 1) and then with DBCO-Oligo (lane 2). (left) Post conjugation clean-up was done using CleanAb Kit for rabbit IgG (lanes 3-5). Excess oligo was successfully removed as shown by SYBR Gold DNA staining and IgG eluted with high recovery (~70%) at neutral pH (lane 4). Desalting was done with Zeba spin desalting columns (lane 5). Only very little IgG remaining on beads (lane 6) (right) Post conjugation clean-up was done using Protein A beads (lanes 3-5). Excess oligo was successfully removed as shown by SYBR Gold DNA staining but IgG elution with low pH failed (lane 4). Most IgG remains on beads (lane 6)
Rabbit IgG (anti beta-tubulin; 80713-1-RR) was conjugated with NHS-Azide (lane 1) and then with DBCO-Oligo (lane 2). (left) Post conjugation clean-up was done using CleanAb Kit for rabbit IgG (lanes 3-5). Excess oligo was successfully removed as shown by SYBR Gold DNA staining and IgG eluted with high recovery (~70%) at neutral pH (lane 4). Desalting was done with Zeba spin desalting columns (lane 5). Only very little IgG remaining on beads (lane 6) (right) Post conjugation clean-up was done using Protein A beads (lanes 3-5). Excess oligo was successfully removed as shown by SYBR Gold DNA staining but IgG elution with low pH failed (lane 4). Most IgG remains on beads (lane 6)
(left) Mouse IgG (anti-Lamin B1 IgG1; 66095-1-Ig, lane 1) was bound to CleanAb rabbit beads (flowthrough in lane 2), then functionalized on beads with NHS-Azide (flowthrough in lane 3) and then with DBCO-Oligo (flowthrough in lane 4). Excess oligo was successfully removed as shown in SYBR Gold DNA staining and the oligo-conjugated IgG was eluted with high recovery at neutral pH (lane 6). Desalting was done with Zeba spin desalting columns (lane 8). Only very little IgG remaining on beads (lane 9). Note: the CleanAb Elution Buffer can interfere with SDS gels, which can make the CleanAb elution appear washed out. (right) Mouse IgG (anti-Lamin B1 IgG1; 66095-1-Ig, lane 1) was bound to Protein A beads (flowthrough in lane 2), Excess oligo was successfully removed as shown in SYBR Gold DNA staining but the oligo-conjugated IgG failed to elute at low pH (lane 6). Most IgG remains on beads (lane 9).
(left) Rabbit IgG (Isotype control, lane 1) was bound to CleanAb rabbit beads (flowthrough in lane 2), then functionalized on beads with NHS-Azide (flowthrough in lane 3) and then with DBCO-Oligo (flowthrough in lane 4). Excess oligo was successfully removed as shown in SYBR Gold DNA staining and the oligo-conjugated IgG was eluted with high recovery at neutral pH (lane 6). Desalting was done with Zeba spin desalting columns (lane 8). Only very little IgG remaining on beads (lane 9). Note: the CleanAb Elution Buffer can interfere with SDS gels, which can make the CleanAb elution appear washed out. (right) Rabbit IgG (Isotype control, lane 1) was bound to Protein A beads (flowthrough in lane 2), Excess oligo was successfully removed as shown in SYBR Gold DNA staining but the oligo-conjugated IgG failed to elute at low pH (lane 6). Most IgG remains on beads (lane 9).
CleanAb vs Protein A
两种方法均可有效去除多余的DNA oligo。但CleanAb试剂盒在结合亲和力、洗脱条件及回收率三项关键指标上均优于Protein A试剂盒，可在简化操作流程的同时减少样品降解，最终获得更高质量的实验结果。
结合力更强
相较于Protein A beads，CleanAb Beads展现出更为优异的亲和力和结合力，穿流液中未检测到偶联IgG即可佐证该优势。CleanAb Beads对各类IgG亚型均具备均一稳定的亲和力，结合力的增强不仅可有效提高产量，还能提高实验重复性，保障纯化结果的稳定性与一致性。
温和天然洗脱
与Protein A beads需采用酸性洗脱条件、可能对抗体结构完整性造成损伤不同，CleanAb Beads可在pH 7.5的温和条件下实现高效洗脱。该温和且接近生理环境的洗脱体系能够有效保护蛋白空间构象与生物活性，特别适用于对敏感的寡核苷酸偶联物、酶学检测实验及下游功能研究等场景。同时，减少样品在低pH环境下的暴露，可最大程度缓解蛋白聚集、变性及目标分子损失等问题。
回收率高
与Protein A beads相比，CleanAb Beads的回收率高达60 - 90%，显著降低了产物损失。其优化后的bead特质和洗脱形式确保了样本的高效洗脱，同时残留的样本极低。相比之下，Protein A常出现洗脱不完全的情况，导致珍贵样品残留在珠子上。使用CleanAb Beads可获得纯度更高的样品、更优的得率，以及更稳定可靠的下游实验表现。
脱盐处理
CleanAb试剂盒可高效去除体系中多余的ssDNA oligos，并在温和天然pH 7.5条件下进行洗脱，有效保持抗体活性和偶联物的结构完整。
缓冲液置换的适用场景
CleanAb洗脱缓冲液中含有的部分组分可能会干扰PAGE检测或影响浓度测量结果。若下游应用需对样品进行精确定量，建议进行缓冲液置换。此外，考虑到偶联抗体在洗脱缓冲液中的长期稳定性因抗体类型而异，同样推荐更换缓冲液后再进行长期保存；并选择在- 80℃条件下储存。
缓冲液置换虽可能造成少量样品损失，但所得产物仍可保持完整的功能，右侧免疫荧光对比结果已对此予以验证。可根据实验需求调整最终稀释比，以获得最佳使用效果。
注：试剂盒不含缓冲液置换层析柱。
Immunostaining of HeLa cells with mouse anti-B23/NPM1 antibody (60096-1-Ig, 1:100, magenta), chemically conjugated to a 15-nt DNA oligonucleotide and purified using CleanAb beads. Detection was performed by hybridization with an imaging DNA strand labeled with ATTO643. Nuclei were counterstained with DAPI (yellow). Direct comparison of CleanAb product and desalted product demonstrates functionality of both products.
常见问题（FAQ）
在哪里可以获取CleanAb试剂盒寡核苷酸去除或染料去除的详细实验方案？
琼脂糖试剂盒与磁珠琼脂糖试剂盒对应的实验方案均可在产品详情页面查阅。
兔IgG专用CleanAb磁珠与小鼠IgG专用CleanAb磁珠是否可结合所有兔源或小鼠源抗体？
兔IgG专用CleanAb磁珠与小鼠IgG专用CleanAb磁珠均采用κ轻链特异性配体，无法结合λ轻链型抗体。绝大多数（＞95%）小鼠及兔IgG抗体为κ轻链亚型，λ轻链抗体仅占极少比例。因此，CleanAb磁珠可适配绝大多数兔源、小鼠源IgG抗体。若不清楚目标抗体的轻链亚型，建议查阅抗体供应商提供的产品资料，或在实验前联系我们技术支持团队。
流穿液中出现偶联后IgG，是什么原因？
NHS酯化偶联后，若偶联IgG结合效果不佳，可调整目标标记分子与IgG的摩尔比。过度标记，即寡核苷酸或染料投料过量，会封闭Fab结合蛋白的结合位点，进而降低结合效率。
重要提示：开展偶联反应前，务必测定IgG实际浓度。供应商标注的抗体浓度不一定完全准确，该操作可有效避免IgG发生过度标记。
需要使用多少磁珠？
实验方案中给出了对应抗体投入量的磁珠使用体积，该体积可按比例线性放大。若处理的抗体量更少，可等比例降低磁珠用量，但抗体回收率会随之下降。
CleanAb试剂盒兼容哪些偶联化学反应？
CleanAb试剂盒适配主流偶联化学方法制备的抗体偶联物，包括NHS酯-氨基偶联、马来酰亚胺-巯基偶联以及点击化学反应。
CleanAb试剂盒仅去除游离寡核苷酸与染料，还是同样可以去除未偶联抗体？
磁珠可直接结合抗体分子，因此会同时捕获偶联抗体与未偶联抗体；游离寡核苷酸、染料及其他非结合组分将在洗涤步骤中被去除。
CleanAb试剂盒是否可用于纳米抗体或抗体片段的纯化？
CleanAb磁珠以物种特异性方式识别抗体Fab区。基于该结合机制，试剂盒可用于木瓜蛋白酶、胃蛋白酶酶切得到的Fab、F(ab')2抗体片段的纯化。
本试剂盒不适用于羊驼来源VHH等纳米抗体片段的纯化。
CleanAb试剂盒能否去除过量染料或体系中其他杂质缓冲液组分？
可以。抗体小分子染料偶联完成后，可使用CleanAb试剂盒进行纯化，同时可去除BSA、Tris、叠氮化钠等缓冲液杂质组分。
洗脱后产物的纯度如何？
经HPLC验证，CleanAb试剂盒可去除95%以上的过量染料，游离寡核苷酸去除率接近100%。洗脱产物中若仍残留微量染料，可通过额外的缓冲液置换步骤进一步去除。
CleanAb试剂盒是否适用于大规模、高通量抗体偶联物纯化工作流程？
适用。CleanAb试剂盒可支持大规模及高通量抗体偶联物纯化，如需定制化解决方案，请联系我们。
相较于体积排阻色谱法与离心过滤法，CleanAb试剂盒具备哪些优势？
离心过滤会造成抗体损失，且往往需要多轮洗涤才能有效去除未反应寡核苷酸；体积排阻色谱会稀释样品，同时需要方法优化才能获得理想的回收率与分离效果，且并非所有实验室均配备色谱设备。CleanAb试剂盒操作流程简便，可实现高且稳定的抗体回收率，高效去除过量寡核苷酸或染料。
CleanAb试剂盒应选择偶联后纯化，还是on-beads后偶联纯化？
CleanAb试剂盒同时支持抗体偶联后纯化与on-beads后偶联纯化两种工作模式，两种方案均可获得稳定、可重复的实验结果。磁珠上原位偶联的核心优势为：抗体先被磁珠捕获，可在偶联反应前去除Tris、BSA、甘氨酸及其他含氨基干扰组分，降低其对偶联反应的干扰，省去偶联前的预处理步骤。
针对NHS酯类偶联反应，优先推荐采用偶联后纯化方案。原因是NHS酯同样可与磁珠固定配体上的官能团发生反应，消耗部分标记试剂，降低偶联效率。若仍需对NHS化学采用磁珠上原位偶联，可适当提高寡核苷酸或染料投料量以补偿该损耗。
