EnzCheck® Direct Phospholipase C Assay kit

特長

• PC-PLC活性を測定する効果的で簡便な方法です。
• 非放射性で安全です。
• 洗浄ステップが不要で、基質を添加して測定できます。迅速です。
• 選択的に基質と反応して明るい緑蛍光（励起/蛍光＝509nm/516nm）を発します。
• キットはPC-PLC抑制をキャラクタライズし、直接に測定をするので複合スクリーンの偽陽性の可能性は排除されます。
• PC-PLCは癌やHIV1-7に関連するアポトーシスや細胞サバイバル等、セルシグナリング・パスウェイにとって重要な役割をしています。
• 96ウェルに200µｌ使用し10マイクロプレート分です。

スペクトルとアッセイデータ
	スペクトルデータ Normalized excitation and emission spectra for PLC substrate after reaction with PC-PLC. Samples were incubated in the presence of 2 U/mL PC-PLC from B. cereus for approximately 60 minutes at room temperature.
PC-PLC検出データ
Triplicate samples from B. cereus were assayed atconcentration of 7.8 mU/mL to 500 mU/mL per well in the presence of 1X PLC substrate and 200 µM lecithin in 1X PLC reaction buffer. Reactions were incubated at room temperature for 60 minutes and fluorescence was measured at Ex/Em 490/520 nm. The inset represents a separate experiment and illustrates the linearity of fluorescence response at low levels of PC-PLC. The average variation of replicates (CV) was less than 3%. Background fluorescence has been subtracted.	IC50 determination for D609, a known competitive PC-PLC inhibitor8-11 with a Ki of 6.4 µM12, using the EnzChek® Direct PLC Assay Kit. Triplicate samples of D609 were titrated in the presence of 1X PLC substrate, 200 µM lecithin and, 100 mU/mL PC-PLC in 1X PLC reaction buffer. Reactions were incubated for 60 minutes at room temperature and fluorescence was measured at Ex/Em 490/520 nm using a fluorescent microplate reader.
	感度と検出範囲データ Sensitivity and dynamic range of the EnzChek® Direct PLC Assay Kit using low-volume 384-well white microplates. Triplicate PC-PLC samples from B. cereus were assayed at concentrations of 31.25 mU/mL to 2,000 mU/mL per well in the presence of 1X PLC substrate and 200 µM lecithin in 1X reaction buffer. Reactions were incubated at room temperature for 60 minutes and fluorescence was measured at Ex/Em 490/520 nm. The average variation of replicates (CV) was lessthan 5%. Background fluorescence has been subtracted.

製品内容

構成品	反応数/キット
	E10215 E10216 96ウェル 200 µｌ/ウェル 200反応分 1,000反応分 384ウェル 20 µｌ/ウェル 2,000反応分 10,000反応分

• 製品のマニュアルはコチラ (182KB)
• PLA2検出用：EnzCheck® Direct Phospholipase A2 Assay kitの製品ページはコチラ
• PLA2新基質：Red/Green BODIPY® PC-Aの製品ページはコチラ

製品情報

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