组织和培养细胞溶酶体富集试剂盒
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组织和培养细胞溶酶体富集试剂盒
Thermo Scientific™

组织和培养细胞溶酶体富集试剂盒

Thermo Scientific™ 组织和培养细胞溶酶体富集试剂盒可从粗细胞和组织提取物中分离和富集完整溶酶体。该试剂盒提供足以制备 25 份提取物的试剂,并使用 OptiPrep 细胞分离基质从污染的细胞结构中对溶酶体进行基于密度的分离。分离的溶酶体可用于多种下游应用,包括用于蛋白质组学研究的 2D/MS了解更多信息
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货号数量
89839230 mL
货号 89839
价格(CNY)
5,955.00
Each
添加至购物车
数量:
230 mL
请求批量或定制报价
价格(CNY)
5,955.00
Each
添加至购物车
Thermo Scientific™ 组织和培养细胞溶酶体富集试剂盒可从粗细胞和组织提取物中分离和富集完整溶酶体。该试剂盒提供足以制备 25 份提取物的试剂,并使用 OptiPrep 细胞分离基质从污染的细胞结构中对溶酶体进行基于密度的分离。分离的溶酶体可用于多种下游应用,包括用于蛋白质组学研究的 2D/MS、电子显微镜检查、疾病谱分析和基因表达、信号转导以及相互作用或定位。

组织和培养细胞溶酶体富集试剂盒的特点:

高效且易于使用—试剂盒试剂和梯度离心操作可从污染结构中分离细胞器
兼容—制备样品用于下游应用,包括 2D/MS、电子显微镜检查、疾病谱分析、基因表达、信号转导和相互作用或定位研究
全套—试剂盒包含足以用于 25 次应用的材料

该试剂盒可从细胞和组织中富集完整细胞器。每个试剂盒使用密度梯度离心操作从杂质细胞结构中分离各种细胞器官。分离出的细胞器可用于多种下游应用,包括 2D/MS、电子显微镜检查、疾病谱分析、基因表达、信号转导和相互作用或定位研究。

组织中的细胞器富集:
处死前,对平均体重为 160 g 的 6 至 8 周龄雌性 Sprague-Dawley 大鼠整夜禁食。切除肝脏和肾脏,用冰冷的 PBS 洗涤,使用手术剪刀进行切碎,然后在冷冰试剂 A 中使用 Polytron™ Tissue Tearer 在 8000 rpm 下进行 45 秒均质处理。随后加入试剂 B,对匀浆离心以去除细胞碎片。将所得的上清液覆盖到不连续梯度的 OptiPrep™ 细胞分离基质上,并离心以分离和富集靶标细胞器。从梯度中取出靶标条带,并采用 Western 印迹进行分析。

细胞中的细胞器富集:
检查了多种细胞系的溶酶体分离:A431 [美国典型培养物保藏中心(ATCC™ 资源中心),#CRL-1555]、HeLa (ATCC, #CCL-2) 和 HepG2 细胞 (ATCC, #HB-8065)。细胞生长至 80-90% 融合。收获并处理约 50-200 mg 湿细胞团用于溶酶体富集。将冰冷的溶酶体试剂 A 添加到细胞中,并使用 Misonix 超声波仪 3000(15 次脉冲,提供 15 W 功率)进行裂解。

随后,添加溶酶体试剂 B,并对匀浆进行离心以去除细胞碎片。将所得的上清液覆盖到不连续梯度的 OptiPrep 细胞分离基质上,并离心以分离和富集靶标细胞器。从梯度中取出靶标条带,连同总裂解物按蛋白量进行标准化(除非另有说明),并采用 Western 印迹进行分析。

相关产品
培养细胞的线粒体分离试剂盒
组织的线粒体分离试剂盒
仅供科研使用。不可用于诊断程序。
规格
数量230 mL
试剂类型细胞器分离、亚细胞分离
足够用于25 份样品,每份含 50-200 mg 细胞或组织
形式液体
产品类型溶酶体富集试剂盒
Unit SizeEach
内容与储存
接收后,在 4°C 下储存。产品在环境温度下运输

常见问题解答 (FAQ)

Which specific ultracentrifuge tubes do you recommend using when working with the Lysosome Enrichment Kit for Tissues and Cultured Cells (Cat. No. 89839)?

We recommend using 8 mL open top ultracentrifuge tubes that are compatible with the specific rotor being used in the lab. Specifically, our R&D team has used Thermo Scientific 03020 tubes (https://www.fishersci.com/shop/products/03020/03020) with Thermo Scientific Sorvall MTX 150 Micro-Ultracentrifuge (Cat. No. 46960) and S58-A Fixed Angle Rotor (Cat. No. 45866). For any other ultracentrifuge, the tubes will need to match the rotor. If the rotor does not take 8 mL tubes, the protocol can be scaled to match the tube size.

Find additional tips, troubleshooting help, and resources within our Protein Dialysis, Desalting, and Concentration Support Center.

Can we use frozen cells and tissues with the Lysosome Enrichment Kit for Tissues and Cultured Cells (Cat. No. 89839)?

We have not tested the isolation of lysosomes from frozen tissues using this kit. We recommend using fresh tissues only as freeze-thaw compromises membrane integrity and it may affect the protein yield.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Can you provide the shelf-life for the Lysosome Enrichment Kit for Tissues and Cultured Cells?

The Lysosome Enrichment Kit for Tissues and Cultured Cells is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended. Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Can I use the Lysosome Enrichment Kit for Tissue and Cultured Cells (Cat. No. 89839) for isolation of the cytosolic fraction?

Lysosome Enrichment Kit for Tissues and Cultured Cells has been designed for isolation and enrichment of intact lysomes from crude cell and tissue extracts. We do not have any guidelines for isolation of the cytosolic fraction as the rest of the density gradient was not defined in-house.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

What kits do you offer for cell fractionation?

We offer the following kits for cell fractionation:

Mem-PER Plus Membrane Protein Extraction Kit, Cat. No. 89842
NE-PER Nuclear and Cytoplasmic Extraction Reagents, Cat. No. 78835
Subcellular Protein Fractionation Kit for Cultured Cells, Cat. No. 78840
Subcellular Protein Fractionation Kit for Tissues, Cat. No. 87790
Mitochondrial Isolation Kit for Cultured Cells, Cat. No. 89874
Mitochondrial Isolation kit for Tissue, Cat. No. 89801
Lysosome Enrichment Kit for Tissues and Cultured Cells, Cat. No. 89839

Find additional tips, troubleshooting help, and resources within our Cell Lysis and Fractionation Support Center.