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[供應]091000449-Accutase™ 胰蛋白酶替代品 091000449
Accutase替換胰酶/EDTA/細胞消化液 091000449
ACCUTASETM細胞消化液 091000449
傳統貼壁細胞消化(胰酶/EDTA)的*替換產品 091000449
MP Biomedicals( MPbio)中國代理重慶市華雅干細胞技術有限公司
Accutase替換胰酶/EDTA/細胞消化液
ACCUTASETM細胞消化液
傳統貼壁細胞消化(胰酶/EDTA)的*替換產品
Accutase替換胰酶/EDTA/細胞消化液描述:
Mpbio公司提供的ACCUTASE™具有蛋白酶和膠原酶活性,是胰酶/EDTA的替換產品,用于從常規組織培養器皿和粘附培養器皿中消化細胞。另外進行細胞計數、轉染或分析檢測(如流式細胞術)前,常加入一定量的accutase,將細胞集落 (clumps)分散成單個細胞然后進行下游實驗。Accutase因其消化方式溫和,對細胞傷害極低,不含任何動物或細菌來源組分等特點成功替代傳統消化產品,廣泛應用于常規貼壁細胞(血清培養細胞和無血清培養細胞)的消化,以及成為干細胞(如hESCs、mESCs等)傳代培養的產品之一。即用型的無菌產品,使用非常方便。
ACCUTASETM細胞消化液配方:
Accutase具蛋白酶和膠原酶活性,以Dulbecco’s PBS溶液(含0.5 mM EDTA·4Na和酚紅)形式供應。
ACCUTASETM細胞消化液優 勢:
● 應用于絕大多數原代細胞和哺乳動物細胞系的消化
實驗驗證的細胞系(cellline):成纖維細胞,角蛋白細胞,血管內皮細胞,肝細胞,血管平滑肌細胞,肝細胞祖細胞,原代雞胚神經細胞,骨髓干細胞,貼壁CHO和BHK細胞,巨噬細,293細胞,L929 細胞,*性小鼠睪丸生殖細胞,3T3,Vero ,COS,HeLa,NT2,MG63, M24,A375轉移性黑色素瘤,神經膠質瘤U251和D54,HT1080纖維肉瘤細胞,Sf9 昆蟲細胞。(參考文獻1-9)
● 溫和有效的消化胚胎干細胞(hESCs和mESCs),凍存后細胞具更高復蘇率(圖2,文獻10)
● 幾分鐘內實現粘附細胞的分離,不需清洗或中和反應,節省細胞傳代時間
● 溫和而快速的分離貼壁細胞,不會破壞流式細胞術檢測的表面抗原
● 溫和消化維持zui高細胞活力,即使長時間消化(45 min)細胞活力達97 ± 3%(圖1)
● 同時結合膠原酶和蛋白酶活性,不含哺乳動物、細菌來源產品或昆蟲桿狀病毒表達系統組分
應 用:
(1)替換胰酶/EDTA(Replacing Trypsin/EDTA)
Accutase的用量和方法不需做改動,特別適用于在無血清和無蛋白培養基等不含血清的培養基內貼壁細胞的分離。相比于胰酶/EDTA,優點在于:即用型,使用方便;大大降低對細胞的傷害;提高細胞產量及細胞活力;增高鋪板效率;改善細胞形態(定性)和細胞生長特性。
(2)功能分析(functional assays)
細胞表面標志物分析,流式細胞術,血清饑餓法檢測細胞靜止狀態,細胞增殖,轉染實驗,細胞趨觸分析,細胞、神經嵴遷移實驗,腫瘤細胞遷移。
(3)組織解離(tissue disaggregation)
在4-25℃下進行組織解離,因酶的特性切忌在37℃操作。根據組織類型和解離溫度優化解離時間。一般情況下,4℃過一夜孵育(12-16hrs),25℃孵育1-2 hrs。
應用實例:
操作步驟:(快速簡單)
(1)37℃或室溫溶解Accutase,用無菌PBS溶液清洗細胞培養容器(培養板、培養瓶等) 。
(2)以無菌操作的方式按 75 cm2表面積加入10 ml Accutase的量覆蓋貼壁細胞;重新放入37℃培養箱內,消化15-20 min。(消化時間可自行調整。大部分細胞處于Accutase中高達1小時,對細胞無影響)
(3)消化結束后,依常規操作進行細胞計數和傳代:不需另做清洗和酶活終止步驟。
訂購信息:
品 牌 | 貨 號 | 名 稱 | 規 格 | 目錄價 | 保存(效期) |
Mpbio | 091000449 | AccutaseTM 細胞分離液 | 100 ml | 486.00 | -20°C(2年) |
應用文獻:
(1)A cell-detachment solution can reduce background staining in theELISPOT assay, Grant A, et al., Methods of Molecular Biology, 2005;302:87-94.
(2)Canine hemangiosarcoma originates from hematopoietic precursors withpotential for endothelial differentiation, Lamerota-Kozicki et al.,Experimental Hematology, Vol. 34 Pages 870-878, April 2006.
(3)The JAK3 inhibitor WHI-P154 prevents PDGF-evoked process outgrowthin human neural precursor cells, Richards et al., Journal ofNeurochemistry, Vol. 97 Page 201, April 2006.
(4)Nuclear factor-КB controls the reaggregation of 3D neurospherecultures in vitro, Widera et al.,European Cells and Materials, Vol. 11,Pages 76-85, 2006.
(5)Autologous adult rodent neural progenitor cell transplantationrepresents a feasible strategy to promote structural repair in thechronically injured spinal cord, Pfeifer, Future Medicine, Pages255-266, July 2006.
(6)Integrin Signalling Regulates the Nuclear Localization and Functionof the Lysophosphatidic Acid Receptor-1 (LPA1) In MammalianCells, Waters et al. Biochemical Journal Immediate Publication, May 2006
(7)Minute numbers of contaminant CD8+ T cells or CD11b+CD11c+ NK cellsare the source of IFN-γ in IL-12/IL-18-stimulated mouse macrophagepopulations, Schleicher et al., Blood Vol. 105,Pages 1319-1328, February 2005.
(8)Cell Permeable Peptide of JNK Inhibitor Prevents Islet ApoptosisImmediay After Isolation and Improves Islet GraftFunction, Noguchi et al., American Journal of Transplantation, Vol.5,Pages 1848-1855, August 2005.
(9) Rescue Purification Maximizes the Use of Human Islet Preparationsfor Transplantation, Ichii et al., American Journal of Transplantation,Vol. 5, Pages 21-30, © 2005.
(10)Efficient Propagation of Single Cells Accutase-Dissociated HumanEmbryonic Stem Cells. RUCHI BAJPAI,MOLECULAR REPRODUCTION AND DEVELOPMENT
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