組成50%グリセロール(v/v)、20 mM NaCl、2 mM MgCl2、20 mMトリス、pH 8.0に調製された、250 U/uLの純度99%以上の酵素
数量5 kU
試薬タイプ細胞溶解用酵素
対応可能対象200 mLライセート
形状液体
製品ラインPierce
製品タイプCell Lysis Enzyme
Unit Size5 kU
組成および保存条件
直射日光を避け、涼しく乾燥した換気の良い場所に保存。
よくあるご質問(FAQ)
At which concentration/dilution should the Pierce Universal Nuclease for Cell Lysis (Cat. No. 88700) be used, and what is the suggested incubation time and incubation temperature?
We would suggest the following procedure (note that you should determine yourself empirically the optimal incubation time, incubation temperature and also dilution):
Please add 25 U or 0.1 µL Pierce Universal Nuclease for Cell Lysis (Cat. No. 88700) to 1 mL of cell lysate (dilution 1:10,000) and incubate at room temperature for 10-15 minutes. If you want to dilute the Nuclease for facilitating pipetting, please do this with Tris- or PBS-buffer at pH 7-8.
After cell lysis, my mass spectrometry sample is very viscous and difficult to pipette. How do I reduce the sample viscosity?
High sample viscosity after lysis is due to release of DNA from the nucleus. Sonication or addition of a nuclease such as the Pierce Universal Nuclease (Cat. No. 88700, 88701, or 88702) can be used to degrade DNA and reduce sample viscosity.
Can you explain how the B-PER Bacterial Protein Extraction Reagent lyses cells?
The B-PER Reagent solution contains a proprietary, mild, non-ionic detergent in 20 mM Tris-HCl, pH 7.5. It effectively disrupts cells and solubilizes native or recombinant proteins without denaturation. The reagent creates holes in the cell membrane that will leak out cytosolic proteins. The sample may become very viscous when the bacterial chromosome is released. We recommend adding DNAse I (Cat. No. 90083) to the reagent to reduce viscosity. For better lysis efficiency and if there are inclusion bodies, we recommend adding Lysozyme (Cat. No. 90082) to the reagent. Alternatively, you may purchase the B-PER Bacterial Protein Extraction Reagent with Enzymes Kit (Cat. No. 90078 or 90079) that includes the B-PER Bacterial Protein Extraction Reagent, DNase I, and Lysozyme.
p53 is active in murine stem cells and alters the transcriptome in a manner that is reminiscent of mutant p53.
Authors:Yan H, Solozobova V, Zhang P, Armant O, Kuehl B, Brenner-Weiss G, Blattner C
Journal:
PubMed ID:25719246
Since it was found that p53 is highly expressed in murine embryonic stem cells, it remained a mystery whether p53 is active in this cell type. We show that a significant part of p53 is localised in the nucleus of murine embryonic stem cells and that the majority of this ... More