Subcellular Protein Fractionation Kit for Cultured Cells
Subcellular Protein Fractionation Kit for Cultured Cells
Thermo Scientific™

Subcellular Protein Fractionation Kit for Cultured Cells

培養細胞用Thermo Scientific細胞内タンパク質分画キットは、5つの異なる細胞コンパートメントからタンパク質を分離および濃縮できます。培養細胞用細胞内タンパク質分画キットの特長:•簡潔 — 機能的な細胞質、膜、核可溶性、クロマチン結合、細胞骨格のタンパク質画分を3時間以内に抽出できます•詳細を見る
製品番号(カタログ番号)数量
788401キット
製品番号(カタログ番号) 78840
価格(JPY)
128,100
Each
お問い合わせください ›
数量:
1キット
一括またはカスタム形式をリクエストする
培養細胞用Thermo Scientific細胞内タンパク質分画キットは、5つの異なる細胞コンパートメントからタンパク質を分離および濃縮できます。

培養細胞用細胞内タンパク質分画キットの特長:

簡潔 — 機能的な細胞質、膜、核可溶性、クロマチン結合、細胞骨格のタンパク質画分を3時間以内に抽出できます
便利 — グラジエントでの超遠心分離を使用しない簡単な手順を実行します
完全 — 単一のキットで、細胞質、膜、可溶性核、クロマチン結合および細胞骨格タンパク質画分を取得できます
培養細胞 — 培養哺乳類細胞で使用するために開発および最適化。組織サンプルには、組織用細胞内分画キットを使用できます
適応 — タンパク質アッセイ、ウェスタンブロッティング、タンパク質アッセイ、電気泳動移動度シフトアッセイ、レポーター遺伝子および酵素活性アッセイなどのダウンストリームアプリケーションに抽出物を使用します。

細胞内分画のアプリケーション:
• 異なる細胞内コンパートメントからのタンパク質の抽出と濃縮
• 同じ細胞から細胞質、核、膜、細胞骨格のタンパク質を分離する
• 細胞質、核、膜、細胞骨格からのタンパク質の細胞のプロファイリング
• タンパク質転座の研究

細胞内タンパク質の分画は、タンパク質局在試験を可能にし、特異的細胞成分からタンパク質を単離します。Thermo Scientific細胞内タンパク質分画キットには、3時間以内で、細胞を機能性細胞質タンパク質、膜タンパク質、水溶性核タンパク質、クロマチン結合タンパク質、細胞骨格タンパク質のそれぞれに段階的に分画し、溶解するための試薬の組み合わせが含まれます。各細胞成分からの抽出物の分画間において、一般的に15%未満の汚染が確認されますが、これはタンパク質の局在および再分布の研究実験にとって十分な純度を保証しています。

培養細胞用Subcellular Protein Fractionation Kitには、4種類の抽出バッファー液、安定化ヌクレアーゼ、およびThermo Scientific停止プロテアーゼ阻害剤カクテルが含まれます。各キットには、それぞれ20µLで50個の細胞ペレットを分画するのに十分な試薬が含まれており、約2 gの細胞ペーストに相当します。(組織のサンプルについては、当社の細胞内タンパク質分画キット(組織用)をご利用ください。)

細胞ペレットに添加する最初のキットの試薬は、細胞膜の透過性を高めて水溶性の細胞質タンパク質を抽出します。二番目の試薬は、血漿、ミトコンドリア、ERゴルジ体膜を抽出しますが、核膜は溶解させません。核を遠心により回収したあと、3番目の試薬により可溶性核抽出物が生じます。ミクロコッカスヌクレアーゼを使用して追加の核抽出を行うと、核タンパク質が放出されます。回収した不溶性ペレットは最後の試薬により抽出され、細胞骨格タンパク質が分離されます。

関連製品
Subcellular Protein Fractionation Kit for Tissues
卓上遠心分離機
研究用途にのみご使用ください。診断目的には使用できません。
仕様
バッファーLysis Buffer
数量1キット
試薬タイプ細胞溶解バッファー、界面活性剤溶液、細胞内分画
対応可能対象50個のサンプル
形状キット、液体
製品タイプSubcellular Protein Fractionation Kit
Unit SizeEach
組成および保存条件
納入後は、キットは-20°Cで保管してください。または上記のように個々のコンポーネントを保管してください。キットはドライアイス入りで出荷されます。

よくあるご質問(FAQ)

What products are available for the isolation of subcellular components?

You can choose from several types of Dynabeads magnetic beads for coating with antibody against the subcellular component of choice. We currently do not offer any ready-to-use Dynabeads magnetic beads against a particular subcellular component, and recommend that you choose the beads that work best with your antibody and your application.

The following groups of Dynabeads magnetic beads can be used for this purpose:

(1) Surface Activated Dynabeads magnetic beads
Antibodies from any origin can be conjugated directly to these uncoated beads through covalent bonds. For better orientation of primary antibody on the beads, a secondary antibody (linker antibody) can be coated on the beads first. The Surface Activated Dynabeads magnetic beads are provided in different sizes (4.5, 2.8 and 1 micron) and the larger ones are mostly preferred for this application.

(2) Secondary Coated Dynabeads magnetic beads
These Dynabeads magnetic beads are pre-coated with a secondary antibody and depending on their specificity, they can be easily coated with your primary antibody within a short time period. These beads are provided in two sizes, 4.5 and 2.8 micron. In addition to these beads, our Dynabeads magnetic beads Protein A or Protein G can also be easily coated with antibody from several species. Please see the following links for information about all available Secondary coated Dynabeads magnetic beads and Dynabeads magnetic beads Protein A and Protein G.

(3) Streptavidin Coated Dynabeads magnetic beads
These Dynabeads magnetic beads are coated with streptavidin and require a biotinylated antibody. These beads are provided in two different sizes, 2.8 and 1 micron.

Find more information about Dynabeads magnetic beads at http://www.thermofisher.com/Dynabeads magnetic beads.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Can you provide the shelf-life for the Subcellular Protein Fractionation Kit for Cultured Cells?

The Subcellular Protein Fractionation Kit for 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 Subcellular Protein Fractionation Kit for Cultured Cells (Cat. No. 78840) for subsequent co-immunoprecipitation (co-IP) analysis of different nuclear proteins?

Yes, the nuclear fraction is compatible with co-immunoprecipitation (co-IP) after dialysis, desalting, or 50:50 dilution. The cytoplasmic and membrane fractions are compatible with co-IP directly after extraction and the pellet fraction is compatible with co-IP after detergent removal.

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

Will M-PER Mammalian Protein Extraction Reagent extract membrane or cytoskeletal proteins?

M-PER Mammalian Protein Extraction Reagent can extract some membrane or cytoskeletal proteins, but the extraction efficiency is not consistent. The reagent was not intended to specifically extract these proteins. We recommend using Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842) for membrane protein extraction or Subcellular Protein Fractionation Kit for Cultured Cells (Cat. No. 78840) for compartmental extraction.

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

引用および参考文献 (3)

引用および参考文献
Abstract
DRIM modulates Src activation and regulates angiogenic functions in vascular endothelial cells.
Authors:Tong J,Dong X,Martin TA,Yang Y,Dong B,Jiang WG
Journal:Cell biology international
PubMed ID:39648301
Downregulated in Metastasis Protein (DRIM) was discovered in malignant epithelial cells and was thought to be mainly a nucleus protein affecting cancer cells. Recent single-cell sequencing analysis suggests that DRIM is abundantly expressed in vascular endothelial cells. There has been no knowledge of the role of DRIM in the endothelium. ... More
Protocol to identify human subcellular alternative protein interactions using cross-linking mass spectrometry.
Authors:Garcia-Del Rio DF,Fournier I,Cardon T,Salzet M
Journal:STAR protocols
PubMed ID:37384523
Since the start of mass-spectrometry-based proteomics, proteins from non-referenced open reading frames or alternative proteins (AltProts) have been overlooked. Here, we present a protocol to identify human subcellular AltProt and decipher some interactions using cross-linking mass spectrometry. We describe steps for cell culture, in cellulo cross-link, subcellular extraction, and sequential digestion. ... More
Evaluation of a Low-Toxicity PARP Inhibitor as a Neuroprotective Agent for Parkinson's Disease.
Authors:Puentes LN,Lengyel-Zhand Z,Reilly SW,Mach RH
Journal:Molecular neurobiology
PubMed ID:33788167
Repurposing PARP-1 inhibitors (PARPi) for non-oncological applications offers an attractive therapeutic strategy for pathological conditions characterized by PARP-1 hyperactivity. In the context of Parkinson's disease (PD), PARP-1 hyperactivity has been linked to neuronal death and disease progression. From a therapy perspective, the evaluation of PARPi as neuroprotective agents may offer ... More