Q：该产品有沉淀，可以直接使用吗？

A：5×SDS-PAGE 蛋白上样缓冲液必须完全溶解后再使用。

Q：该产品有沉淀，能直接煮沸溶解吗？

A：不可以。在室温或不超过 37°C 的水浴中溶解 5×SDS-PAGE 上样缓冲液。水浴溶解后立即室温存放，尽量避免长时间置于水浴中。

Q：蛋白自身的电荷会对缓冲液有影响吗？

A：不会的。5×SDS-PAGE 蛋白上样缓冲液中 SDS 可与蛋白质结合使蛋白质－SDS 复合物上带有大量的负电荷，这时蛋白质本身的电荷完全被 SDS 掩盖，因此，电泳速度只是与其分子量大小有关。

Q：上样过程需要添加指示剂吗？

A：需要，一般采用溴酚蓝用作电泳指示剂，可大概指示电泳结束的时间。

1. Zhang W, Lu J, Feng L, et al. Sonic hedgehog-heat shock protein 90β axis promotes the development of nonalcoholic steatohepatitis in mice. Nat Commun. 2024;15(1):1280. Published 2024 Feb 12. doi:10.1038/s41467-024-45520-8(IF:16.6)
2. Xie S, Sun W, Zhang C, et al. Metabolic Control by Heat Stress Determining Cell Fate to Ferroptosis for Effective Cancer Therapy. ACS Nano. 2021;15(4):7179-7194. doi:10.1021/acsnano.1c00380(IF:15.881)
3. Wang Y, Chen J, Tian J, et al. Tryptophan-sorbitol based carbon quantum dots for theranostics against hepatocellular carcinoma. J Nanobiotechnology. 2022;20(1):78. Published 2022 Feb 14. doi:10.1186/s12951-022-01275-2(IF:10.435)
4. Pan S, Pei L, Zhang A, et al. Passion fruit-like exosome-PMA/Au-BSA@Ce6 nanovehicles for real-time fluorescence imaging and enhanced targeted photodynamic therapy with deep penetration and superior retention behavior in tumor. Biomaterials. 2020;230:119606. doi:10.1016/j.biomaterials.2019.119606(IF:10.273)
5. Rui W, Xiao H, Fan Y, et al. Systemic inflammasome activation and pyroptosis associate with the progression of amnestic mild cognitive impairment and Alzheimer's disease. J Neuroinflammation. 2021;18(1):280. Published 2021 Dec 2. doi:10.1186/s12974-021-02329-2(IF:8.322)
6. Chao H, Zheng L, Hsu P, et al. IL-13RA2 downregulation in fibroblasts promotes keloid fibrosis via JAK/STAT6 activation. JCI Insight. 2023;8(6):e157091. Published 2023 Mar 22. doi:10.1172/jci.insight.157091(IF:8.0)
7. Dong B, Wang C, Zhang J, et al. Exosomes from human umbilical cord mesenchymal stem cells attenuate the inflammation of severe steroid-resistant asthma by reshaping macrophage polarization. Stem Cell Res Ther. 2021;12(1):204. Published 2021 Mar 24. doi:10.1186/s13287-021-02244-6(IF:6.832)
8. Jiang M, Tang T, Liang X, et al. Maternal sevoflurane exposure induces temporary defects in interkinetic nuclear migration of radial glial progenitors in the fetal cerebral cortex through the Notch signalling pathway. Cell Prolif. 2021;54(6):e13042. doi:10.1111/cpr.13042(IF:6.831)
9. Cheng GP, Guo SM, Yin Y, Li YY, He X, Zhou LQ. Aberrant Expression of Mitochondrial SAM Transporter SLC25A26 Impairs Oocyte Maturation and Early Development in Mice. Oxid Med Cell Longev. 2022;2022:1681623. Published 2022 Apr 13. doi:10.1155/2022/1681623(IF:6.543)
10. Cao G, Li P, He X, et al. FHL3 Contributes to EMT and Chemotherapy Resistance Through Up-Regulation of Slug and Activation of TGFβ/Smad-Independent Pathways in Gastric Cancer. Front Oncol. 2021;11:649029. Published 2021 Jun 4. doi:10.3389/fonc.2021.649029(IF:6.244)
11. Zhou J, Wu L, Xu P, Li Y, Ji Z, Kang X. Filamin A Is a Potential Driver of Breast Cancer Metastasis via Regulation of MMP-1. Front Oncol. 2022;12:836126. Published 2022 Mar 11. doi:10.3389/fonc.2022.836126(IF:6.244)
12. Wu YL, Xue YR, Guo ZT, et al. Furmonertinib (Alflutinib, AST2818) is a potential positive control drug comparable to rifampin for evaluation of CYP3A4 induction in sandwich-cultured primary human hepatocytes. Acta Pharmacol Sin. 2022;43(3):747-756. doi:10.1038/s41401-021-00692-7(IF:6.150)
13.  Li D, Ding J, Liu TL, et al. SARS-CoV-2 receptor binding domain radio-probe: a non-invasive approach for angiotensin-converting enzyme 2 mapping in mice [published correction appears in Acta Pharmacol Sin. 2022 Feb 3;:]. Acta Pharmacol Sin. 2022;43(7):1749-1757. doi:10.1038/s41401-021-00809-y(IF:6.150)
14. Zhang B, Sun H, Zhao F, et al. Characterization and Genomic Analysis of a Novel Jumbo Bacteriophage vB_StaM_SA1 Infecting Staphylococcus aureus With Two Lysins. Front Microbiol. 2022;13:856473. Published 2022 Apr 28. doi:10.3389/fmicb.2022.856473(IF:5.640)
15. Zhang T, Feng S, Li J, et al. Farnesoid X receptor (FXR) agonists induce hepatocellular apoptosis and impair hepatic functions via FXR/SHP pathway. Arch Toxicol. 2022;96(6):1829-1843. doi:10.1007/s00204-022-03266-6(IF:5.153)
16. Shi X, Huang X, Chen R, et al. The transcribed ultraconserved element uc.51 promotes the proliferation and metastasis of breast cancer by stabilizing NONO. Clin Exp Metastasis. 2021;38(6):551-571. doi:10.1007/s10585-021-10128-5(IF:5.150)
17. Tang B, Sun EZ, Zhang ZL, et al. Sphingomyelin-Sequestered Cholesterol Domain Recruits Formin-Binding Protein 17 for Constricting Clathrin-Coated Pits in Influenza Virus Entry. J Virol. 2022;96(5):e0181321. doi:10.1128/JVI.01813-21(IF:5.103)