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Merck
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重要文件

71296

Millipore

PhosphoSafe蛋白质提取试剂

Novagen′s PhosphoSafe Extraction Buffer efficiently extracts cytosolic proteins from mammalian and insect cells while preserving their phosphorylation state.

同義詞:

Protein Extraction Reagent

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About This Item

分類程式碼代碼:
41106510
NACRES:
NA.77
暫時無法取得訂價和供貨情況

品質等級

100

形狀

liquid

製造商／商標名

Novagen®

儲存條件

OK to freeze

技術

immunoblotting: suitable

應用

sample preparation

運輸包裝

dry ice

儲存溫度

−70°C

相關類別

一般說明





PhosphoSafe提取缓冲液可有效地从哺乳动物和昆虫细胞中提取胞质蛋白，同时保持其磷酸化状态。该试剂含有与CytoBuster蛋白质提取试剂（Trim 2001）相同的配方，但还包括四种磷酸酶抑制剂：氟化钠、钒酸钠、β-甘油磷酸酯和焦磷酸钠。PhosphoSafe提取试剂与激酶测定、蛋白质相互作用分析和其他应用兼容。该试剂以方便的25 mL瓶装提供（71296-3为1瓶，71296-4为5瓶）。

特點和優勢

  • 提取过程中保留的磷酸化状态
  • 与激酶测定相容

警告

毒性：有毒（F）

其他說明

Trim，J.E. and Sawyer，D.L.2001.inNovations12，5.

法律資訊

NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany

儲存類別代碼

12 - Non Combustible Liquids

水污染物質分類（WGK）

WGK 2

閃點（°F）

Not applicable

閃點（°C）

Not applicable

分析證明 (COA)

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Adrienne R Gibson et al.
Cancer research, 80(7), 1401-1413 (2020-02-12)
Pharmacologic ascorbate treatment (P-AscH-, high-dose, intravenous vitamin C) results in a transient short-term increase in the flux of hydrogen peroxide that is preferentially cytotoxic to cancer cells versus normal cells. This study examines whether an increase in hydrogen peroxide is

Questions

1–2 of 2 Questions  

  1. what is recomended volume for 50mg tissue extraction? would you recomend adding protease ihnibitor too? could you share protocol for protein extraction from liver using this phosphosafe extraction reagent?

    1 answer

    1. A protocol for liver tissue particularly is not offered. Here is a protocol for tissue in general:

      Tissue homogenates are prepared by homogenizing tissue in ice-cold PhosphoSafe extraction buffer using 70 ul buffer/mg tissue. Homogenates are incubated at room temperature for 5 mins, sonicated on ice, and then subjected to centrifugation (spun at 16,000 X g, 5 mins, at 4 Celsius). The resulting supernatant is collected (the end-user will want to keep this, and can discard the insoluble pellet). Finally, protein concentration is determined by a protein assay (e.g., Bradford assay).

      To minimize proteolysis, extraction can be performed on ice, and protease inhibitors can be added.
      Please see the link below to access the full product user guide:
      https://www.emdmillipore.com/product/PhosphoSafe-Extraction-Reagent,EMD_BIO-71296#anchor_USP

      Helpful?

  2. I am doing protein isolation from the organic phase remaining after RNA isolation using Trizol/Chloroform. How can I use PhosphoSafe™ Extraction Reagent in this case?

    1 answer

    1. Phosphosafe has been tested for use in aqueous formats such as cell culture supernatants or lysates. The performance of the product in an organic phase following fractionation with Trizol has not been determined. To discuss further, we kindly ask you to navigate to the link https://www.sigmaaldrich.com/techservice, click on "Product Technical Inquires" under the Products Section with all the required information so that a member of our team can reach out to you to assist further. Thank you.

      Helpful?

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