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- 详细信息
- 用户评价
- 文献和实验
- 技术资料
- 英文名：
Minute™ Plasma Membrane-Derived Lipid Raft Isolation Kit
- 保存条件：
4℃
- 规格：
20 Preps
MinuteTM 质膜脂筏分离试剂盒
目录号：LR-042
描述：
脂筏是含有高水平胆固醇和鞘脂的小质膜结构域。脂筏被发现存在于质膜（PM）和一些内膜系统中，如线粒体相关膜（MAMS）和内质网等。脂筏参与许多细胞过程，如信号转导，膜转运和蛋白质分选。脂质修饰的蛋白质和一些跨膜蛋白质集中在脂筏中，而其他蛋白质被排除在外。脂筏还被发现与质膜上的Na+ / K+ ATP酶相关。传统的脂筏分离方法是从总膜结构中分离出脂筏，而不能分质膜或者细胞器膜上的脂筏。为了克服传统方法的缺点，我们开发了此款基于离心管柱技术的质膜脂筏分离试剂盒。首先较大的质膜囊泡被分离出来，然后使用含有非离子表面活性剂缓冲液处理，通过台式离心机即可离心分离质膜脂筏部分。不使用传统的匀浆机和超速离心机，1h左右即可高度富集质膜脂筏。
*如需分离总的脂筏，请使用 Minute TM 脂筏分离试剂盒 Cat#LR-039
应用：
提取的蛋白可用于SDS-PAGE ，ELISA ，IP，2D胶分析，质谱，酶活检测以及其他应用。
试剂盒组分(20T):
1. Buffer A 15ml
2. Buffer B 10ml
3. Buffer C 10ml
4. 研磨棒 2 个
5. 离心管柱+收集管 20 套
重要产品信息:
1.所有离心步骤应在 4℃ 室温下或冷冻离心机中进行。
2.研究蛋白质磷酸化，在使用前应在缓冲液 A 中加入磷酸酶抑制剂。如果蛋白质降解可能会导致下游实验问题，在使用前加入蛋白酶抑制剂在缓冲 A 和 B 中。（各类抑制剂的添加方法请按照抑制剂母液比例，例如母液是 100x，添加时按照 1：100 添加，即 1ml 缓冲液中添加 10ul 抑制剂）。
3.推荐使用 BCA 试剂盒测定蛋白质浓度。
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文献和实验参考文献：
1.Bu, Y., Teng, Q., Feng, D., Sun, L., Xue, J., & Zhang, G. (2021). YLMY Tyrosine Residue within the Cytoplasmic Tail of Newcastle Disease Virus Fusion Protein Regulates Its Surface Expression to Modulate Viral Budding and Pathogenicity. Microbiology Spectrum, 9(3), e02173-21.
2.Rashkovan, M., Albero, R., Gianni, F., Perez-Duran, P., Miller, H. I., Mackey, A. L., ... & Ferrando, A. A. (2021). Intracellular cholesterol pools regulate oncogenic signaling and epigenetic circuitries in Early T-cell Precursor Acute Lymphoblastic Leukemia. Cancer discovery.
3.Shan-Shan Gao; Run Shi; Jing Sun; Yanhong Tang; Jing-Feng Li; et al(2022).GPI Anchored Ligand-BioID2-Tagging System Identifies Galectin-1 Mediating Zika Virus Entry.SSRN Electronic Journal.
4.Fiore, D., Proto, M. C., Franceschelli, S., Pascale, M., Bifulco, M., & Gazzerro, P. (2022). In Vitro Evidence of Statins’ Protective Role against COVID-19 Hallmarks. Biomedicines, 10(9), 2123.
5.Lim, J. H., Ahmad, K., Chun, H. J., Hwang, Y. C., Qadri, A. F., Ali, S., ... & Lee, E. J. (2022). IgLON4 Regulates Myogenesis via Promoting Cell Adhesion and Maintaining Myotube Orientation. Cells, 11(20), 3265.
6.Sha, Y. L., Liu, Y., Yang, J. X., Wang, Y. Y., Gong, B. C., Jin, Y., ... & Zhao, Q. (2022). B3GALT4 remodels the tumor microenvironment through GD2-mediated lipid raft formation and the c-met/AKT/mTOR/IRF-1 axis in neuroblastoma. Journal of Experimental & Clinical Cancer Research, 41(1), 1-21.
7.Gao, S.-S., Shi, R., Sun, J., Tang, Y., Zheng, Z., Li, J.-F., Li, H., Zhang, J.,Leng, Q., Xu, J., Chen, X., Zhao, J., Sy, M.-S., Feng, L., Li, C., GPI-Anchored Ligand-BioID2-Tagging System Identifies Galectin-1 Mediating Zika Virus Entry, ISCIENCE (2022), doi: 10.1016/j.isci.2022.105481.
8.Li, B., Ding, Z., Calbay, O. et al. FAP is critical for ovarian cancer cell survival by sustaining NF-κB activation through recruitment of PRKDC in lipid rafts. Cancer Gene Ther (2022). doi.org/10.1038/s41417-022-00575-x
9.Oh, S., Jo, S., Bajzikova, M., Kim, H. S., Dao, T. T., Rohlena, J., ... & Park, S. (2023). Non-bioenergetic roles of mitochondrial GPD2 promote tumor progression. Theranostics, 13(2), 438.
10.Zhao, Q., Su, H., Jiang, W., Luo, H., Pan, L., Liu, Y., ... & Tan, B. (2023). IGF-1 Combined with OPN Promotes Neuronal Axon Growth in Vitro Through the IGF-1R/Akt/mTOR Signaling Pathway in Lipid Rafts. Neurochemical Research, 1-12.
11.Mirza, I., Haloul, M., Hassan, C., Masrur, M., Mostafa, A., Bianco, F. M., ... & Mahmoud, A. M. (2023). Adiposomes from Obese-Diabetic Individuals Promote Endothelial Dysfunction and Loss of Surface Caveolae. Cells, 12(20), 2453.
12.Khalifa, B. A., Guijarro, A., Ravera, S., Bertola, N., Adorni, M. P., Papotti, B., ... & Nencioni, A. (2023). Cyclic fasting bolsters cholesterol biosynthesis inhibitors’ anticancer activity. Nature Communications, 14(1), 6951.
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14. Piergiorgio La Rosa; Jessica Tiberi; Enrico Palermo; Roberta Stefanelli; Sofia Maria Luigia Tiano; et al.(2024).The inactivation of the Niemann Pick C1 cholesterol transporter restricts SARS-CoV-2 entry into host cells by decreasing ACE2 abundance at the plasma membrane. Cell & Bioscience. DOI 10.1186/s13578-024-01331-4
15.Anqi Zhou; Bao Xue; Jiayi Zhong; Junjun Liu; Ran Peng; et al.(2025). Cholesterol-rich lipid rafts mediate endocytosis as a common pathway for respiratory syncytial virus entry into different host cells.Microbiology Spectrum.DOI 10.1128/spectrum.01192-25
KSUD14-STS ASSAY FOR TAGGING Lr21
New D14 primers. Lr21L: CGC TTT TAC CGA GAT TGG TC Lr21R: TCT GGT ATC TCA CGA AGC CTT Lr21 : 669 bp lr21 -Fielder: 757 bp lr21 -Wichita: 774 bp Recipe (in 25 µl reaction
Inducible shRNA Lentiviral Vectors
TE Buffer, pH 8.0 to 8 µl 5 µl 2) Remove the LR Clonase™ II enzyme mix from -20° C and thaw on ice (~ 2 minutes). 3) Vortex the LR Clonase™ II enzyme mix
柱式法 Minute TM 胞质胞核分离试剂盒操作方法：A． 悬浮细胞样品（包括植物，细菌，酵母和真菌制备的原生质体）1.500Xg，3 分钟低速离心收集细胞，用预冷的 PBS 清洗一次2. 将细胞转移到 1.5 ml 离心管中，3000 rpm 离心 1-5 分钟，弃去上清。3. 按表格 1 加入适量的胞浆提取缓冲液（请注意样品及裂解液的比例，以达到最佳效果）, 涡旋大力震荡 15 秒, 冰上孵育 5 分钟, 混匀. 接转胞质胞核分离步骤。B. 贴壁细胞1. 贴壁细胞生长至融合度 90
