Silver:LysateforWestern
Grow cellsHarvestSpin downWash with PBSSpin down enough cells to collect a 50-100 μL cell pellet in a FastPrep tubePrepare lysis buffer (beforehand) and chill on icePMSF: 1 mL per 100 mL bufferPLAAC: 100 μL per 100 mL buffer0.5 M Benzamidine: 260 μL per 100 mL bufferUse fresh PMSF: dissolve 0.035g per 1mL 100%EtOH2.5 mM MgCl23 mM KCl0.5% Triton X-100in PBSPBSMT:Add protease inhibitorsAdd 50-100 μL of cold lysis buffe......閱讀全文
Silver:－Lysate－for－Western
Grow cellsHarvestSpin downWash with PBSSpin down enough cells to collect a 50-100 μL cell pellet in a FastPrep tubePrepare lysis buffer (beforehand) a
Studier－Lysate－Prep
SummaryHow to make a lysate from a plaque preparation. We also use this protocol for preparation of a quick stock from previously made lysate prep.Pro
Silver－Enhancement－...
實驗概要The method provides a silver enhancement protocol for immunoassay.主要試劑Prepare the following reagents fresh daily except for the citrate buffer.1.
Silver－Staining－Protocol
1x 40min - overnight?????50% MeOH, 12% Acetic Acid1x 30min??????????????????????????????????50% MeOH, 12% Acetic Acid, 0.05% 37% Formaldehyde3x 20min?
Silver:－Colony－PCR
Zymolyase Solution:Regular stock of zymolyase is 10 mg/ml diluted in water, mix by inverting, not all will go into solution (filter it) store aliquots
美國實驗室wetern方法
WESTERN BLOT PROTOCOLIn a Western blot, proteins that are separated on polyacrylamide gels on the basis of size are transferred to a membrane for dete
Silver－Acetate－Autometallography－(AMG)
In the early eighties, a series of papers were published by Gorm DANSCHER, Aarhus, Denmark, to introduce a reliable and easy-to-handle technique for l
SSR－GEL－and－Silver－Staining－Protocol
I.?EQUIPMENT:DNA sequencing unit (35 x 45 cm) & 2000V power supplyClampsLg. plastic trays (4), about 43 x 50 x 8 cm, and one lidTwo rocking platformsH
Silver:－TimeLapse－Microscopy
Pad Preparation1. Microwave 2% agarose (mix of low-melt and normal, to taste) in Thorn media (see below). (If you have used different percentages of a
SDI檢測儀XFsilver
SDI檢測儀/污染指數儀/污染指數測定儀 型號：XF-silver 指數（SDI）值，也稱之為FI（Fouling Index）值，是水質指標的重要參數之一。它代表了水中顆粒、膠體和其他能阻塞各種水凈化設備的物體含量。通過測定SDI值，可以選定相應的水凈化技術或設備。 在反滲透水處
SDI檢測儀XFsilver
SDI檢測儀/污染指數儀/污染指數測定儀 型號：XF-silver 指數（SDI）值，也稱之為FI（Fouling Index）值，是水質指標的重要參數之一。它代表了水中顆粒、膠體和其他能阻塞各種水凈化設備的物體含量。通過測定SDI值，可以選定相應的水凈化技術或設備。 在反滲透水處理
銀染(silver－staining)操作規程
實驗原理：在堿性條件下，用甲醛將蛋白帶上的硝酸銀（銀離子）還原成金屬銀，以使銀顆粒沉積在蛋白帶上。染色的程度與蛋白中的一些特殊的基團有關，不含或者很少含半胱氨酸殘基的蛋白質有時候呈負染。銀染的詳細機制還不是非常清楚。 試劑：乙醇、冰醋酸、乙酸鈉、硫代硫酸鈉、硝酸銀、碳酸鈉、甘氨酸或EDTA.Na
FlagHA－double－tag－IP
實驗概要Isolate the unknown protein that could interact with bait from cell lysate, for potential Mass spectrometry.主要試劑50mM Tris, pH8.020 mM glycerol b-p
蛋白質檢測
·?????????Protein detection?(Aberdeen's Lab)The method used to locate the proteins following 2D-PAGE depends on the nature of the original sample.
Embryo－Lysates－－Immunoprecipitation
Embryo lysatesTake 25 embryos and place into 1.7ml centrifuge tube.Rinse once in lysis buffer (add ~ 1ml) and remove by aspirationAdd 500 μL lysis buf
VSVG標簽融合蛋白檢測，封閉實驗手冊
VSV-G,來源于水泡性口炎病毒的融合性外殼G糖蛋白，常被用于逆轉錄病毒和慢病毒載體的生物醫學研究。VSV-G標簽通常融合于目的蛋白的N-或者C-端，以便使用免疫組化方法來進行觀察和分析。Fig.1.Immunofluorescence staining (1:1,000) of VSV-G fus
低背景的蛋白質銀染(silver－staining)方法
我們做蛋白質電泳的人都知道，銀染色很靈敏，有很強的說服力，但通常膠背景較深，不易掃描。在這里介紹一下低背景的蛋白質銀染方法。Step Reagent Time/minFix 50%EtOH, 12%HAC, 0.1%HCHO, 38%H2O 60Rinse 50% EtOH 5 min, 3 tim
Cell－and－tissue－lysis－hub
This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison
Myc標簽融合蛋白檢測，封閉應用數據
Myc 標簽融合蛋白?- 檢測，封閉Myc Tag來源于c-myc基因表達產物，其序列為EQKLISEEDL。以高親和力聞名的Myc Tag的單克隆抗體（克隆號2D5），可以檢測天然和變性的Myc融 合蛋白，被廣泛用于WB、IF、IP實驗。在天然洗脫條件下，使用Myc標簽多 肽來與重組蛋白進
Western雜交
實驗概要本實驗介紹了Western雜交的基本流程。主要試劑液氮，提取緩沖液(1 x PBS，10ug/mL ?Leupeptin，1 mM PMSF，5 mMBenzamidine. 2mM EDTA)，轉移緩沖液(39mM Glycine，48mM ?Tris，0.037% SDS，20%甲醇)，
Western雜交
實驗概要本實驗介紹了Western雜交的基本流程。主要試劑液氮，提取緩沖液(1 x PBS，10ug/mL ?Leupeptin，1 mM PMSF，5 mMBenzamidine. 2mM EDTA)，轉移緩沖液(39mM Glycine，48mM ?Tris，0.037% SDS，20%甲醇)，
Western－Blotting
實驗概要Western Blot (AP)主要試劑1. Membrane Blocking buffer:5% Milk ? 0.05% of Tween 20 PBS2. antibody dilution buffer: PBS 0.05% of Tween20 1.0% Milk(or BSA
Western－雜交
Western?雜交（主要內容如下）Preparing of Protein LysatesWestern BlottingFar Western BlottingSemi Dry BlottingStripping MembranesTrouble Shooting and OthersPrepa
Western雜交
Western雜交l??????組織印跡的Western雜交在硝酸纖維素膜上制備組織印跡1．在塑料板上放置兩層Whatman 1號濾紙，在濾紙上方放上一張普通紙，然后鋪上一層硝酸纖維素膜。2．用雙面刀片從植物組織如大豆的莖上切取一塊切片（厚度約為1mm）。如果組織表面是濕的，則在Kimwipes紙上
Western－Blotting
1. Optional: "Renature" gel- this is thought to permit some refolding of proteins and may be important in finding epitope recognition of monoclonal an
Western－blotting樣品準備－（一）
實驗概要Preparation of ?lysis buffers, protease and phosphatase inhibitors, lysate from cell ?culture, lysate from tissues, protein concentration, samples
Immunoprecipitation...－（二）
3. ImmunoprecipitationImmunoprecipitation can be performed using antibodies by different ?methods. The use of these methods is based on the requiremen
Western實驗步驟
一、樣品制備?對于Western Blotting實驗而言，樣品處理是關鍵步驟之一，獲得的蛋白樣品必須均質、可溶，并解離成單個多肽亞基，且盡量減少相互間的聚集，使其最終僅依賴本身的分子量大小進行分離。當目標蛋白的含量很低時，需要選取目標蛋白含量較高的組織或細胞器（如核抽提物），或者通過免疫沉淀等方式
Western－Blotting－Protocol
實驗概要The western blot ?(sometimes called the protein immunoblot) is a widely used analytical ?technique used to detect specific proteins in the given s
western－blot原理
Western Blot法采用的是聚丙烯酰胺凝膠電泳，被檢測物是蛋白質，經過聚丙烯酰胺凝膠電泳分離的蛋白質樣品，轉移到固相載體上，且能保持電泳分離的多肽類型及其生物學活性不變。以固相載體上的蛋白質或多肽作為抗原，與對應的抗體起免疫反應，再與酶或同位素標記的第二抗體起反應，經過底物顯色或放射自顯影以檢