CleanAb Oligo清除试剂盒
快速和简便地从Oligo偶联抗体中去除多余的寡核苷酸
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适用于Rabbit IgG或Mouse IgG（所有亚型）
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高效去除游离Oligo
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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 Oligo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 rabbit anti-CD147 antibodies (11989-1-AP, 1:100, orange hot), 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 (cyan). Direct comparison of CleanAb product and desalted product demonstrates functionality of both products. While signal intensity is lower for the desalted sample, successful staining can still be achieved.
