Description
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Whole Cell Lysate, 1 mg (2 mg/ml)
General Notes:
Growth media: DMEM supplemented with 10% FBS
Cell lysate was prepared by homogenization in RIPA buffer (25mM Tris HCl pH 7.6, 150mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS) supplemented with 5 µg/ml of aprotinin and 5 µg/ml of leupeptin. Cell debris were removed by centrifugation (5 minutes at 12,000 RPM). Concentration of total protein was determined using BCA protein assay.
Storage Instructions
Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
Volume
0.5 ml
Concentration
1 mg at 2 mg/ml
Purity
Whole Cell Lysate
Altogen Labs Research Services:
Altogen Labs provides GLP-compliant contract research studies for pre-clinical research, IND applications, and drug development. Biology CRO services include: Xenograft models (30+), development of stable cell lines, ELISA assay development, cell-based and tissue targeted RNAi studies, safety pharm/tox assays, and other studies (visit AltogenLabs.com).
Volume Options:
- 1 mg of A204 Whole Cell Lysate (Catalog #8890)
- 1 mg of A375 Whole Cell Lysate (Catalog #8891)
- 1 mg of A431 Whole Cell Lysate (Catalog #8892)
- 1 mg of A549 Whole Cell Lysate (Catalog #8893)
- 1 mg of AGS Whole Cell Lysate (Catalog #8894)
- 1 mg of AR42J Whole Cell Lysate (Catalog #8895)
- 1 mg of ARPE-19 Whole Cell Lysate (Catalog #8896)
- 1 mg of ASMC Whole Cell Lysate (Catalog #8897)
- 1 mg of AsPC-1 Whole Cell Lysate (Catalog #8898)
- 1 mg of Astrocyte Whole Cell Lysate (Catalog #8899)
- 1 mg of B16-F10 Whole Cell Lysate (Catalog #8900)
- 1 mg of BALB/3T3 Whole Cell Lysate (Catalog #8901)
- 1 mg of bEnd-3 Whole Cell Lysate (Catalog #8902)
- 1 mg of Beta-TC-6 Whole Cell Lysate (Catalog #8903)
- 1 mg of BHK-21 Whole Cell Lysate (Catalog #8904)
- 1 mg of BJ Whole Cell Lysate (Catalog #8905)
- 1 mg of BMS2 Whole Cell Lysate (Catalog #8906)
- 1 mg of BT-20 Whole Cell Lysate (Catalog #8907)
- 1 mg of BxPC-3 Whole Cell Lysate (Catalog #8908)
- 1 mg of C2C12 Whole Cell Lysate (Catalog #8909)
- 1 mg of C6 Whole Cell Lysate (Catalog #8910)
- 1 mg of Caco-2 Whole Cell Lysate (Catalog #8911)
- 1 mg of Caki-1 Whole Cell Lysate (Catalog #8912)
- 1 mg of Calu-3 Whole Cell Lysate (Catalog #8913)
- 1 mg of Calu-6 Whole Cell Lysate (Catalog #8914)
- 1 mg of Capan-1 Whole Cell Lysate (Catalog #8915)
- 1 mg of CFPEo Whole Cell Lysate (Catalog #8916)
- 1 mg of CHO Whole Cell Lysate (Catalog #8917)
- 1 mg of CHO-K1 Whole Cell Lysate (Catalog #8918)
- 1 mg of Chromaffin Whole Cell Lysate (Catalog #8919)
- 1 mg of CLBPEC Whole Cell Lysate (Catalog #8920)
- 1 mg of COLO205 Whole Cell Lysate (Catalog #8921)
- 1 mg of COS-7 Whole Cell Lysate (Catalog #8922)
- 1 mg of CT26.WT Whole Cell Lysate (Catalog #8923)
- 1 mg of D-407 Whole Cell Lysate (Catalog #8924)
- 1 mg of D-451 Whole Cell Lysate (Catalog #8925)
- 1 mg of Detroit551 Whole Cell Lysate (Catalog #8926)
- 1 mg of DI-TNC1 Whole Cell Lysate (Catalog #8927)
- 1 mg of DLD-1 Whole Cell Lysate (Catalog #8928)
- 1 mg of DU145 Whole Cell Lysate (Catalog #8929)
- 1 mg of FaDu Whole Cell Lysate (Catalog #8930)
- 1 mg of Fibroblast Whole Cell Lysate (Catalog #8931)
- 1 mg of H9c2 Whole Cell Lysate (Catalog #8932)
- 1 mg of HCAEC Whole Cell Lysate (Catalog #8933)
- 1 mg of HCC1937 Whole Cell Lysate (Catalog #8934)
- 1 mg of HCN-1A Whole Cell Lysate (Catalog #8935)
- 1 mg of HCS-2 Whole Cell Lysate (Catalog #8936)
- 1 mg of HCT-116 Whole Cell Lysate (Catalog #8937)
- 1 mg of HEK-293 Whole Cell Lysate (Catalog #8938)
- 1 mg of HeLa Whole Cell Lysate (Catalog #8939)
- 1 mg of HeLa-S3 Whole Cell Lysate (Catalog #8940)
- 1 mg of Hep-3B Whole Cell Lysate (Catalog #8941)
- 1 mg of Hepa 1-6 Whole Cell Lysate (Catalog #8942)
- 1 mg of HepG2 Whole Cell Lysate (Catalog #8943)
- 1 mg of HMEC Whole Cell Lysate (Catalog #8944)
- 1 mg of HT-1080 Whole Cell Lysate (Catalog #8945)
- 1 mg of HT-29 Whole Cell Lysate (Catalog #8946)
- 1 mg of HUH-7 Whole Cell Lysate (Catalog #8947)
- 1 mg of HUVEC Whole Cell Lysate (Catalog #8948)
- 1 mg of IMR-90 Whole Cell Lysate (Catalog #8949)
- 1 mg of IRR-MRC5 Whole Cell Lysate (Catalog #8950)
- 1 mg of KB Whole Cell Lysate (Catalog #8952)
- 1 mg of Keratinocyte Whole Cell Lysate (Catalog #8953)
- 1 mg of L6 Whole Cell Lysate (Catalog #8954)
- 1 mg of LNCAP Whole Cell Lysate (Catalog #8955)
- 1 mg of LoVo Whole Cell Lysate (Catalog #8956)
- 1 mg of LS-174T Whole Cell Lysate (Catalog #8957)
- 1 mg of MCF-7 Whole Cell Lysate (Catalog #8958)
- 1 mg of MDA-MB Whole Cell Lysate (Catalog #8959)
- 1 mg of MDCK Whole Cell Lysate (Catalog #8960)
- 1 mg of MEF Whole Cell Lysate (Catalog #8961)
- 1 mg of MG-63 Whole Cell Lysate (Catalog #8962)
- 1 mg of MIA PaCa Whole Cell Lysate (Catalog #8963)
- 1 mg of MRC5 Whole Cell Lysate (Catalog #8964)
- 1 mg of NCI-H1299 Whole Cell Lysate (Catalog #8965)
- 1 mg of NCI-H1975 Whole Cell Lysate (Catalog #8966)
- 1 mg of NCI-H23 Whole Cell Lysate (Catalog #8967)
- 1 mg of NCI-H292 Whole Cell Lysate (Catalog #8968)
- 1 mg of NCI-H358 Whole Cell Lysate (Catalog #8969)
- 1 mg of NCI-H441 Whole Cell Lysate (Catalog #8970)
- 1 mg of NCI-H460 Whole Cell Lysate (Catalog #8971)
- 1 mg of NCI-N87 Whole Cell Lysate (Catalog #8972)
- 1 mg of Neuro-2a Whole Cell Lysate (Catalog #8973)
- 1 mg of NIH3T3 Whole Cell Lysate (Catalog #8974)
- 1 mg of NTERA-2 Whole Cell Lysate (Catalog #8975)
- 1 mg of PANC-1 Whole Cell Lysate (Catalog #8976)
- 1 mg of PC-12 Whole Cell Lysate (Catalog #8977)
- 1 mg of PC-3 Whole Cell Lysate (Catalog #8978)
- 1 mg of PGHAM Whole Cell Lysate (Catalog #8979)
- 1 mg of RD Whole Cell Lysate (Catalog #8980)
- 1 mg of SK-BR3 Whole Cell Lysate (Catalog #8981)
- 1 mg of SK-MEL-28 Whole Cell Lysate (Catalog #8982)
- 1 mg of SKNAS Whole Cell Lysate (Catalog #8983)
- 1 mg of SK-N-MC Whole Cell Lysate (Catalog #8984)
- 1 mg of SK-N-SH Whole Cell Lysate (Catalog #8985)
- 1 mg of SK-OV-3 Whole Cell Lysate (Catalog #8986)
- 1 mg of SW480 Whole Cell Lysate (Catalog #8987)
- 1 mg of T98G Whole Cell Lysate (Catalog #8988)
- 1 mg of U87 Whole Cell Lysate (Catalog #8989)
- 1 mg of VERO Whole Cell Lysate (Catalog #8990)
- 1 mg of Weri-Rb-1 Whole Cell Lysate (Catalog #8991)
- 1 mg of ZR-75-1 Whole Cell Lysate (Catalog #8992)
- 1 mg of 293 Whole Cell Lysate (Catalog #8993)
- 1 mg of 293T/17 Whole Cell Lysate (Catalog #8994)
- 1 mg of 3LL Whole Cell Lysate (Catalog #8995)
- 1 mg of 3T3 L1 Whole Cell Lysate (Catalog #8996)
- 1 mg of 4T1 Whole Cell Lysate (Catalog #8997)
- 1 mg of 786-O Whole Cell Lysate (Catalog #8998)
AltogenBiosystems是一家开发和制造用于生命科学研究,药物发现和开发的转染试剂盒的生物技术公司。转染试剂盒针对特定癌细胞系和原代细胞培养进行了优化,可将生物分子有效递送到靶组织中。通过先进的试剂配方和优化的转染方案实现体外(癌细胞系)和体内(动物组织靶向试剂、癌细胞系)递送货物分子,包括质粒DNA,各种类型的RNA(mRNA,siRNA,shRNA,microRNA),蛋白质和小分子研究。
Altogen生命科学公司致力于研发,生产和销售特定细胞系的转染试剂,用于细胞间生物分子的传递,并通过对转染试剂类型的设计将siRNA和质粒DNA有效地转入不同的细胞系和原代细胞内。Altogen公司开发的聚合物,脂质体,纳米粒子为基础的转染技术分别针对分子生物学,组合化学,和细胞生物学而分别应用。Altogen定制服务提供符合GLP要求定制研究服务,包括代稳定的细胞系,细胞银行和冷冻保存,焦磷酸测序,克隆,RNA干扰(RNAi)和基因沉默服务,发展分析,siRNA文库筛选,并转染服务。稳定的肿瘤细胞株和原代细胞的产生,可以是非常昂贵和费时。该公司的细胞培养科学家的细胞株的选择,无论是利息或shRNA表达载体的稳定表达的基因改造。标准的RNAi技术服务,包括设计与合成的siRNA的利益,验证siRNA的沉默效率,siRNA转染条件的优化,使高效的基因沉默细胞系或原代培养细胞的靶基因。转染培养细胞的瞬时或稳定的引入外源性分子和遗传物质(即RNA或DNA),通常是在生物实验室用来研究基因功能,基因表达的调节,生化映射,突变分析,和蛋白质的生产。科学家利用各种载体分子,这种分子,使质粒DNA(PDNA),信使RNA(mRNA),短干扰RNA(siRNA),小分子RNA(miRNA)的,并进入肿瘤细胞株和原代细胞的蛋白质的基因交付。不幸的是,无单提货的方法或转染试剂,可以适用于所有类型的细胞,细胞的细胞毒性和转染效率显着不同,取决于试剂,协议,并正在利用细胞类型。Altogen生物系统公司提供超过60种类型的细胞的预优化转染试剂盒。纳米粒子,脂质和聚合物基ALTOGEN®在体内转染试剂,使交付功能的RNA和DNA分子在体内。PEG脂质体在体内输送系统减少由于PEG修饰的先天免疫反应,并提供高效的siRNA转染的DNA,并在体内的蛋白质。由科学“杂志(2010年12月17日):PEG脂质体在体内转染试剂盒siRNA的特色Altogen生物系统功能的特定细胞系转染试剂盒
120+细胞转染试剂和活体组织靶向试剂盒制造商AltogenBiosystems是一家生物技术公司,开发和制造用于生命科学研究、药物发现和开发的转染试剂盒。Altogen®体内转染试剂可有效地将生物分子导入靶组织。细胞转染试剂盒针对特定的癌细胞系和原代细胞进行了优化。通过先进的试剂配方和优化的转染方案实现货物分子(DNA、RNA、蛋白质)的高效传递。AltogenBiosystems利用高分子化学、分子和细胞生物学的专业知识,开发了新的体内外给药技术。转染是将外源分子导入培养细胞中,常用于研究基因功能、基因表达调控、生化定位和蛋白质生产。不幸的是,由于细胞毒性和转染效率的差异很大,并且取决于所使用的试剂、方案和细胞类型,因此没有一种单一的传递方法或转染试剂可应用于所有类型的细胞。AltogenBiosystems为120多个癌细胞系和原代细胞类型提供优化的转染试剂盒和电穿孔产品。体内转染试剂可实现组织靶向给药。Altogen的转染试剂盒包括用于体外(癌细胞系)和体内(用于动物研究的组织靶向试剂)转染的转染增强剂试剂和转染复合物冷凝器。Altogen实验室提供符合GLP的实验室合同研究服务。我们的生物CRO服务包括异种移植物的疗效、IND应用的pharm/tox研究和安全性测试、分析开发(ELISA、IC-50、qPCR)、90多个异种移植物动物模型、RNAi和基因沉默服务。Altogen的细胞培养科学家通过在28天内培育出稳定的细胞系,将选择的细胞系转化为稳定表达感兴趣的基因。
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一、机械裂解法主要有以下两中:
1.热休克(Thermal shock),既反复冻溶法,是一种常用的机械裂解方式比,通常由冷冻和解冻两部分组成(freezing and thawing),.原理:由于细胞内冰粒形成和剩余细胞液的盐浓度增高引起溶胀,使细胞结构破碎.冷冻通常在液氮或-20°C冰上进行,解冻可以在37、50、65 或100℃水浴中进热休克比化学裂解温和,但是很有效,有资料表明用热休克和溶菌酶与SDS的方法获得了90%的细胞裂解率.
2.超声波处理(Ultrasonication)既利用超声加热的方法,把细胞破碎.但这种处理会导致DNA 的断裂,所以加热不宜过剧烈,要设定好超声时间和间隙时间,一般超声时间不超过5秒,间隙时间最好大于超声时间,这些都有利于保护酶的活性.bead-beating 也是常用的机械处理方式,有报道指出bead-beating 比热休克和化学裂解的细胞裂解效果更好 虽然DNA产量较高,但通常得到的DNA片段较小.
二、 化学裂解和酶裂解法(在提核酸时联合使用)
主要是裂解液处理法,细胞裂解液的主要目的有以下几种:(1)利用去污剂破坏脂质双分子层,破裂细胞;(2)溶解蛋白;(3)蛋白变性使其稳定;(4)抑制蛋白酶活性.
主要根据不同的目的,裂解液的组成有所不同,主要有提取核酸和蛋白两中.在提取RNA或DNA时,我们主要是要充分裂解细胞,得到更多的核酸;如果我们的目的是蛋白,那要根据蛋白的位置、特性等因素考虑裂解液,在提取蛋白后,再根据实验需要复性蛋白等.以下是细胞裂解中常用试剂和其作用:
50 mM Tris-HCl pH 7.4(缓冲体系),150 mM NaCl(等渗体系),1 mM PMSF (强大的蛋白酶抑制剂),1 mM EDTA(变性剂和稳定剂),5 μg/ml Aprotinin(蛋白酶抑制剂),5 μg/ml Leupeptin(蛋白酶抑制剂),1% Triton x-100(破坏细胞),1% Sodium deoxycholate(中度变性剂和蛋白溶解剂),0.1% SDS(强变性剂和蛋白溶解剂).7M 尿素,2M硫脲(可以提高膜蛋白的融解),蛋白酶K等.
不知单纯使用冰浴中超声裂解够了吗?还是说要选用裂解液呢?最主要的就是我不知道该怎么赔这个裂解液,使其COX活性不产生影响