GSTActivityFluorometricAssay
实验概要The experiment provides a simple, fluorescence-based in vitro assay for detecting the GST activity using a fluorescence plate reader. The assay utilizes monochlorobimane (MCB), a dye that reacts with glutathione. The free form of MCB is almost nonfluorescent, whereas the dye fluoresces blue (ex/em = 380/461 nm) when it reacts with glutathione. GST catalizes the MCB-glutathione ......阅读全文
GST-Activity-Fluorometric-Assay
实验概要 The experiment provides a simple, fluorescence-based in vitro assay for detecting the GST activity using a fluorescence plate reader. The ass
Assay-of-Phospholipase-A-Activity
Phospholipases of the A type constitute a large family of esterases that catalyze the hydrolysis of the fatty acid ester bonds in phospholipids a
Measuring-PLD-Activity-In-Vivo
Phospholipase D (PLD) hydrolyzes structural phospholipids like phosphatidylcholine (PC) and phosphatidylethanolamine (PE) into phosphatidic acid
Regulation-of-transcriptional-activity-by-PML
The PML nuclear bodies are ring-shaped nuclear substructures associated with the regulation of transcription, trans
Mechanism-of-Acetaminophen-Activity-and-Toxicity
Acetaminophen is one of the worlds most commonly used drugs, used for the treatment of pain and fever. Like other NSAIDs (non-steroidal anti-inflammat
Alanine-Transaminase-Activity-Assay
实验概要Alanine Transaminase (ALT) is a transaminase (EC 2.6.1.2) also called serum glutamic pyruvic transaminase (SGPT). Alanine Transaminase is found
GST标签抗体GST标签与IFIP等多种应用
近年来在原核表达体系中,谷胱甘肽S转移酶GST表达纯化系统的应用更为普遍,它的来源是日本血吸虫的25kDa大小的GST蛋白。GST标签系统具有蛋白表达产率高、表达产物纯化方便,以及利于GST抗体制备等特点和优势。GST融合蛋白在水溶液中可溶,可从细菌裂解液中提取,在不变性的条件下通过亲和层析得到。G
Hydrolytic-Activity-of-Bacterial-Supernatants-for-Fungal-Suppression
As the fungal growth suppression by biocontrol agents (BCA) in solid media using dual plate assay has some issues regarding nutrient limitation. A pr
Assay-of-superoxide-dismutase-activity1
Assay of superoxide dismutase activity by combining electrophoresis and densitometryAbstract. A modified technique was developed to assay superoxide d
Assay-of-superoxide-dismutase-activity3
Botanical Bulletin of Academia Sinica, Vol. 37, 1996The method using NBT as a superoxide radical competitor and a color indicator was also explored to
Assay-of-superoxide-dismutase-activity2
Cuvette holders in the sample chamber of the spectrophotometer were thermo-controlled at 25°C. For the blank test, 100 ml of 50 mM potassium phosphate
Purification-of-GST-Fused-Proteins
Day 1Set up an overnight culture in 100 ml LMM broth or 100 ml terrific broth containing 100ul 100 mg/mlAmpDay 2Add 40-50 ml o/n culture to 1 lt terri
GST蛋白纯化步骤
制备细胞裂解物:1.每100ml培养物的细胞沉淀悬于4mlPBS;2.加入溶菌酶至终浓度1mg/ml,冰上放置30min;3.用针筒将10ml 0.2%Triton X-100强行注入细胞裂解物中,剧烈震动数次混匀;4.加入DNase和RNase至终浓度5μg/ml,4℃震动温育10min;5.4℃
GST融合蛋白(GST-fusion-protein-purification)的表达与纯化
原理GST 纯化系统是利用GST (glutathione-S-transferase )融合蛋白与固定的谷胱甘肽(GSH)通过硫键共价亲和,通过GSH交换洗脱的原理来进行纯化 。1ml树脂大约可结合5-8 mg融合蛋白,并可反复使用数次。试剂u IPTG(异丙基硫代-β-D-半乳糖苷) 2
Methods-for-the-Measurement-of-a-Bacterial-Enzyme-Activity-in-Cell-Lysates
Abstract The kinetic characteristics and regulation of aspartate carbamoyltransferase activity were studied in lysates and cell extracts of He
Pelp1-Modulation-of-Estrogen-Receptor-Activity
Proline-,glutamic acid-,leucine-rich protein 1 (Pelp1 also known as:modulator of nongenomic activity of estrogen or MNAR) is a recently discovered tra
Cellfree-System-for-the-examination-of-apoptotic-activity
IntroductionIn our lab we use the term 'cell-free system' when we talk about the examination of apoptotic activity in cytoplasmic extracts. T
什么是水活度(Water-Activity)
水活度被定义为当前可用“自由水”的量,和水分含量没有直接的对比关系,水活度值范围在0(绝对干燥)和1(100%相对湿度)之间。当物质与环境空气发生水分交换活动会在表面形成对微生物生长的理想条件,这样会影响微生物的稳定性。水活度同时也是影响食品中化学反应的重要因素。
GST-融合蛋白沉降实验
实验材料 细胞裂解液其中蛋白质 35S 用标记试剂、试剂盒 裂解缓冲液SDS 聚丙烯酰胺凝胶探针GST 蛋白仪器、耗材 沸水浴翻转祥品旋转仪谷胱甘肽琼脂糖球珠实验步骤 材料缓冲液和溶液将缓冲液稀释到适当浓度。裂解缓冲液20 mmol/LTris-Cl(pH8.0)200 mml/L NaCl1 mm
GSTpulldown操作流程
实验材料 探针蛋白原核蛋白细胞蛋白组织蛋白提取物试剂、试剂盒 NaClKClNa2HPO4KH2PO4Triton-100IPTGPMSF(苯甲基磺酰氟)CocktaierImmobilized Glutathione无水乙醇仪器、耗材 定容瓶锥形瓶高速离心机低温冰箱超声仪EP管层析柜
GSTpulldown操作流程
实验目的:体外检测蛋白质与蛋白质之间相互作用。用于验证两个已知蛋白的相互作用,或者筛选与已知蛋白相互作用的未知蛋白。实验原理:利用重组技术将探针蛋白与GST(Glutathione S transferase)融合,融合蛋白通过GST与固相化在载体上的GTH(Glutathione)亲和结合。因此,
GST融合蛋白的准备
Preparation of Glutathione-S-Transferase (GST) Fusion ProteinsMargret B. Einarson and Elena N. Pugacheva Foxx Chase Cancer Center, Philadelphia, PA 19
GST-融合蛋白沉降实验
GST 沉降实验与 far western(方案 2) 相比有—个优点:探针蛋白是在更自然的环境下与可能的搭档蛋白孵育,因而增强了相互作用的有效性。本实验来源于分子克隆实验指南(第三版)下册,作者:〔美〕J. 萨姆布鲁克 D.W. 拉塞尔。实验材料细胞裂解液其中蛋白质 35S 用标记试剂、试剂盒裂解
GSTpulldown操作流程
实验目的:体外检测蛋白质与蛋白质之间相互作用。用于验证两个已知蛋白的相互作用,或者筛选与已知蛋白相互作用的未知蛋白。 实验原理:利用重组技术将探针蛋白与GST(Glutathione S transferase)融合,融合蛋白通过GST与固相化在载体上的GTH(Glutathione)
GSTpull-down-技术
实验概要本实验详细介绍了GST-pull down 技术的操作流程,主要包括:GST蛋白的大量制备,谷胱甘肽-琼脂糖树脂的处理,GST蛋白的结合,细胞裂解液的预清除及细胞裂解液的探测。实验步骤1. 大量制备GST蛋白 1) 活化冻存菌种GST和GST-X:按1:50比例将表达蛋白的冻存菌液加入5
GSTpulldown操作流程
实验目的:体外检测蛋白质与蛋白质之间相互作用。用于验证两个已知蛋白的相互作用,或者筛选与已知蛋白相互作用的未知蛋白。实验原理:利用重组技术将探针蛋白与GST(Glutathione S transferase)融合,融合蛋白通过GST与固相化在载体上的GTH(Glutathione)亲和结合。因此,
GSTpulldown操作流程
实验概要体外检测蛋白质与蛋白质之间相互作用。用于验证两个已知蛋白的相互作用,或者筛选与已知蛋白相互作用的未知蛋白。实验原理利用重组技术将探针蛋白与GST(Glutathione S transferase)融合,融合蛋白通过GST与固相化在载体上的GTH(Glutathione)亲和结合。因此,当与
酵母GST蛋白纯化方法
GST Fusion Protein Purification from Yeast5 ml overnight culture of your favorite yeast in your favorite medium.Inoculate 50 ml and grow 30o C shaking
GSTpulldown操作流程
利用重组技术将探针蛋白与GST(Glutathione S transferase)融合,融合蛋白通过GST与固相化在载体上的GTH(Glutathione)亲和结合,已广泛应用于分子生物学(1)证实两种蛋白可能有相互作用(2)寻找能与已知蛋白发生相互作用的未知蛋白。实验方法原理利用重组技术将探针蛋
GST-融合蛋白沉降实验
实验材料 细胞裂解液 其中蛋白质 35S 用标记 试剂、试剂盒 裂解缓冲液 SDS 聚丙烯酰胺凝胶