Q：柱子反压过高是什么原因？

A：填料可能被堵塞：裂解液中可能含微小的固体颗粒，建议上柱前使用滤膜（0.22µm 或 0.45µm）过滤，或离心去除。

Q：为什么洗脱组分中无目的蛋白？

A：蛋白可能是包涵体：可通过电泳检测裂解液，分析上清中是否有目的蛋白，包涵体蛋白需按照包涵体蛋白的纯化方式；

Q：洗脱组分不纯（含多种蛋白）是什么原因？

A： 样品中含有其他 His 标签蛋白：通过调节 pH 值或咪唑浓度来优化洗杂条件。再使用其他纯化手段（如去离子交换，疏水等）进一步纯化洗脱组分。

Q：为什么填料颜色会变浅或变成白色？

A：镍离子脱落或者剥离，按照填料再生的操作重新挂镍离子。

Q：产品规格的体积指的是什么？

A：指的是填料的体积，不包括保护液的。

 Q：填料流速慢的原因？

 A：1.柱子在清洗平衡步骤流速减慢，柱子下筛板堵塞或者是试剂中有杂质堵塞。2.上样过程中流速逐渐降低，样品中有不溶物 堵塞填料或者是样品粘稠。少数情况蛋白挂柱后 流速也会减慢。 3.洗杂洗脱步骤流速减慢，蛋白不稳定 在柱子上沉淀，可以选择低温条件纯化，或者是在整个纯化过程中样品试剂中加入甘油保护蛋白。

1. Cui Q, Bi H, Lv Z, et al. Diverse CMT2 neuropathies are linked to aberrant G3BP interactions in stress granules. Cell. 2023;186(4):803-820.e25. doi:10.1016/j.cell.2022.12.046(IF:64.5)
2. Xia B, Shen X, He Y, et al. SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target. Cell Res. 2021;31(8):847-860. doi:10.1038/s41422-021-00519-4(IF:25.617)
3. Deng X, Xiao Y, Tang X, Liu B, Lin H. Arabidopsis α-Aurora kinase plays a role in cytokinesis through regulating MAP65-3 association with microtubules at phragmoplast midzone. Nat Commun. 2024;15(1):3779. Published 2024 May 6. doi:10.1038/s41467-024-48238-9(IF:16.6)
4. Zhao S, Chen Y, Chen F, et al. Sas10 controls ribosome biogenesis by stabilizing Mpp10 and delivering the Mpp10-Imp3-Imp4 complex to nucleolus. Nucleic Acids Res. 2019;47(6):2996-3012. doi:10.1093/nar/gkz105(IF:11.147)
5. Li S, Yang X, Huang H, et al. Arabidopsis ACYL-ACTIVATING ENZYME 9 (AAE9) encoding an isobutyl-CoA synthetase is a key factor connecting branched-chain amino acid catabolism with iso-branched wax biosynthesis. New Phytol. 2022;233(6):2458-2470. doi:10.1111/nph.17941(IF:10.152)
6. Xie L, Yan T, Li L, et al. An HD-ZIP-MYB complex regulates glandular secretory trichome initiation in Artemisia annua. New Phytol. 2021;231(5):2050-2064. doi:10.1111/nph.17514(IF:10.152)
7. Feng Y, Chen JJ, Xie NB, et al. Direct decarboxylation of ten-eleven translocation-produced 5-carboxylcytosine in mammalian genomes forms a new mechanism for active DNA demethylation. Chem Sci. 2021;12(34):11322-11329. Published 2021 Jul 21. doi:10.1039/d1sc02161c(IF:9.825)
8. Li X, Sun D, Feng H, et al. Efficient arsenate reduction in As-hyperaccumulator Pteris vittata are mediated by novel arsenate reductases PvHAC1 and PvHAC2. J Hazard Mater. 2020;399:122895. doi:10.1016/j.jhazmat.2020.122895(IF:9.038)
9. Liu ZH, Xu HL, Han GW, et al. Self-Assembling Nanovaccine Enhances Protective Efficacy Against CSFV in Pigs. Front Immunol. 2021;12:689187. Published 2021 Jul 21. doi:10.3389/fimmu.2021.689187(IF:7.561)
10. Xie L, Yan T, Li L, et al. The WRKY transcription factor AaGSW2 promotes glandular trichome initiation in Artemisia annua. J Exp Bot. 2021;72(5):1691-1701. doi:10.1093/jxb/eraa523(IF:6.992)
11. Shu C, Wang L, Zou C, et al. Function of Foxl2 and Dmrt1 proteins during gonadal differentiation in the olive flounder Paralichthys olivaceus [published online ahead of print, 2022 Jun 16]. Int J Biol Macromol. 2022;215:141-154. doi:10.1016/j.ijbiomac.2022.06.098(IF:6.953)
12. Chen T, Li Y, Xie L, et al. AaWRKY17, a positive regulator of artemisinin biosynthesis, is involved in resistance to Pseudomonas syringae in Artemisia annua. Hortic Res. 2021;8(1):217. Published 2021 Oct 1. doi:10.1038/s41438-021-00652-6(IF:6.793)
13. Lu Y, Liu ZH, Li YX, Xu HL, Fang WH, He F. Targeted Delivery of Nanovaccine to Dendritic Cells via DC-Binding Peptides Induces Potent Antiviral Immunity in vivo. Int J Nanomedicine. 2022;17:1593-1608. Published 2022 Apr 5. doi:10.2147/IJN.S357462(IF:6.400)