RIPA Lysis and Extraction Buffer
RIPA Lysis and Extraction Buffer
Thermo Scientific™

RIPA Lysis and Extraction Buffer

Thermo Scientific RIPA Lysis and Extraction Bufferは培養哺乳類細胞用の高品質ですぐに使用可能な一般的な細胞溶解試薬で、組成が完全に公開されています。RIPAバッファーの特長:•便利—すぐに使用可能な溶液;自身で成分を調製す必要なし• 柔軟—レポーターアッセイ詳細を見る
製品番号(カタログ番号)数量
89901250 mL
89900100 mL
製品番号(カタログ番号) 89901
価格(JPY)
54,000
250 mL
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数量:
250 mL
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Thermo Scientific RIPA Lysis and Extraction Bufferは培養哺乳類細胞用の高品質ですぐに使用可能な一般的な細胞溶解試薬で、組成が完全に公開されています。

RIPAバッファーの特長:

便利—すぐに使用可能な溶液;自身で成分を調製す必要なし
柔軟—レポーターアッセイ、タンパク質アッセイ、イムノアッセイ、およびタンパク質精製など、多くのアプリケーションに適合
汎用—細胞質、膜、および核のタンパク質を抽出可能
公開されている組成—独自の成分を含有していないため、ユーザーは起こり得る適合性の問題を完全にコントロールし知ることが可能

このRIPAバッファーは、プレートで培養した細胞やペレット化した懸濁細胞などの培養哺乳類細胞を効果的に溶解し、タンパク質を抽出します。この一般的な試薬は、膜、核、および細胞質のタンパク質を抽出し、レポーターアッセイ、Thermo Scientific BCA Protein Assay、イムノアッセイ、およびタンパク質精製など、多くのアプリケーションに適合します。Thermo Scientific Halt Protease Inhibitor Cocktail(製品番号78430)やHalt Phosphatase Inhibitor Cocktail(製品番号78420)などの阻害剤もRIPAバッファー組成に適合し、使用前に添加することでタンパク質分解を防ぎ、タンパク質のリン酸化を維持できます。

RIPAバッファーの名前は、開発目的であったオリジナルアプリケーション、すなわち放射性免疫沈降法(radio-immunoprecipitation assay)に由来します。今日、この同位体アッセイ法がラボで実施されることはほとんどありませんが、この溶解バッファー製剤の頭字語は広く使用され続けています。RIPA細胞溶解試薬は3種類の非イオン性およびイオン性の界面活性剤を含有しているため、さまざまな細胞タイプからタンパク質を効果的に抽出します。この界面活性剤の短所は、他の溶解試薬と比較して、特定のダウンストリームアプリケーションとの適合性が相対的に乏しい点です。

関連製品
Pierce™ IP Lysis Buffer
M-PER™ Mammalian Protein Extraction Reagent
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
フォーマット液体
数量250 mL
容量(メートル法)250 mL
製品タイプ抽出バッファー
Unit Size250 mL
組成および保存条件
受け取り後4℃で保存。

よくあるご質問(FAQ)

What are the standard lysis buffers used with mammalian cells for detection of protein expression by immunoprecipitation (IP) or Western blot analysis?

The most commonly used buffer is RIPA Buffer with SDS. We offer RIPA Buffer (Cat. Nos. 89900 and 89901). We also offer the Pierce IP Lysis buffer (Cat. Nos. 87787 and 87788) as well as M-PER (Cat. Nos. 78501, 78503, and 78505).

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

Does RIPA Lysis and Extraction Buffer (Cat. No. 89900, 89901) contain protease or phosphatase inhibitors?

RIPA Lysis and Extraction Buffer (Cat. No. 89900, 89901) does not contain protease or phosphatase inhibitors. If desired, you may add protease and/or phosphatase inhibitors, such as Halt Protease Inhibitor Cocktail (Cat. No. 78410) and Halt Phosphatase Inhibitor Cocktail (Cat. No. 78420) to the RIPA Lysis and Extraction Buffer to prevent proteolysis and maintain phosphorylation status of proteins. We recommend adding protease and phosphatase inhibitors immediately before use.

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

Why is it not recommended that I use RIPA buffer for protein A280 measurements with my NanoDrop spectrophotometer?

RIPA buffer produces a particularly strong absorbance signal at the 280 nm wavelength. As a result, it will either over estimate or under estimate protein concentrations and interfere with the protein purity ratio.
Protein samples in RIPA buffer should be quantified via the Pierce Protein 660 or BCA colorimetric assays using a full spectrum NanoDrop model.

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

Why is there low phosphorylation of the proteins when I use the RIPA Lysis and Extraction Buffer?

Low phosphorylation is usually due to phosphatase activity. We recommend adding a Halt Phosphatase Inhibitor Cocktail to the buffer before use.
Alternatively, the protein is not phosphorylated or phosphorylated at a low level.

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

What if proteolysis occurs when I use RIPA Lysis and Extraction Buffer?

Proteolysis indicates that no protease inhibitors were added. We recommend adding a Halt Protease Inhibitor Cocktail to the RIPA Lysis and Extraction Buffer before use.

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

引用および参考文献 (7)

引用および参考文献
Abstract
Not just fat: investigating the proteome of cetacean blubber tissue.
Authors:Kershaw JL,Botting CH,Brownlow A,Hall AJ
Journal:Conservation physiology
PubMed ID:29479430
Mammalian adipose tissue is increasingly being recognized as an endocrine organ involved in the regulation of a number of metabolic processes and pathways. It responds to signals from different hormone systems and the central nervous system, and expresses a variety of protein factors with important paracrine and endocrine functions. This ... More
Comparison of methods to isolate proteins from extracellular vesicles for mass spectrometry-based proteomic analyses.
Authors:Subedi P,Schneider M,Philipp J,Azimzadeh O,Metzger F,Moertl S,Atkinson MJ,Tapio S
Journal:Analytical biochemistry
PubMed ID:31401005
Extracellular vesicles (EVs) are cell-derived membrane-bound organelles that have generated interest as they reflect the physiological condition of their source. Mass spectrometric (MS) analyses of protein cargo of EVs may lead to the discovery of biomarkers for diseases. However, for a comprehensive MS-based proteomics analysis, an optimal lysis of the ... More
Alternative direct-to-amplification cell lysis techniques for forensically relevant non-sperm cells.
Authors:Arya R,Hudson BC,Green TD
Journal:Journal of forensic sciences
PubMed ID:37779342
While efforts have been made to reduce the pervasive backlog of sexual assault evidence collection kits, the actual laboratory process remains very time-consuming due to the requirement of a differential lysis step before DNA purification, as well as intricate mixture analysis towards the end of the DNA workflow. Recently, an ... More
TGFB1-induced extracellular expression of TGFBIp and inhibition of TGFBIp expression by RNA interference in a human corneal epithelial cell line.
Authors:Yellore VS, Rayner SA, Aldave AJ
Journal:Invest Ophthalmol Vis Sci
PubMed ID:20881301
'To report the increased production of extracellular transforming growth factor ß-induced protein (TGFBIp) by human corneal epithelial cells (HCECs) after induction by TGFB1 and the inhibition of TGFBIp production in induced and noninduced HCECs by RNA interference (RNAi).' ... More
Repair of full-thickness femoral condyle cartilage defects using allogeneic synovial cell-engineered tissue constructs.
Authors:Pei M, He F, Boyce BM, Kish VL
Journal:Osteoarthritis Cartilage
PubMed ID:19128988
Synovium-derived stem cells (SDSCs) have proven to be superior in cartilage regeneration compared with other sources of mesenchymal stem cells. We hypothesized that conventionally passaged SDSCs can be engineered in vitro into cartilage tissue constructs and the engineered premature tissue can be implanted to repair allogeneic full-thickness femoral condyle cartilage ... More