Phosphate(Sodium)bufferChart
Phosphate (Sodium) buffer ChartStock solution A2 M monobasic sodium phosphate, monohydrate (276g/L)Stock solution B2 M dibasic sodium phosphate (284 g/L).Mixing an appropriate volume (ml) of A and B as shown in the table below and diluting to a total volume of 200 ml, a 1 M phosphate buffer of the required pH at room temperature. A B pH 92.0 8.0 0.890.0 10.0 5.987.7 12.3 6.085.......阅读全文
Human－Placental－Alkaline－Phosphatase－Stain
The following protocol is for detection of human placental alkaline phosphatase (AP) in cultured cells. Human placental AP is heat stable, unlike othe
戊二醛固定细胞【Harvard－University】
Yu-Li Wang Lab,University of massachusetts Medical School(UMASS)http://ylwang.umassmed.edu/protocol/cfs/glutar.htmMaterials
细胞沉淀可以用蛋白loading－buffer溶解吗
细胞沉淀里面是可以用这个溶解的，只不过在溶解的时候它的速度可能会更快的。
Tris缓冲液（TrisHCl－buffer）的配制
将121g的Tris碱溶解于约0.9L水中，再根据所要求的pH（25℃下）加一定量的浓盐酸（11.6N），用水调整终体积至1L。浓盐酸的体积（ml）pH8.6142128.53846566671.3769.08.88.68.48.28.07.87.67.47.2
红细胞裂解液（RBC－Lysis－Buffer,10×）－操作步骤
、组织细胞样本的常规操作1、制备细胞悬液: 新鲜组织经过胰蛋白酶或胶原酶等消化处理，通过适当方法制备成细胞悬液，离心弃上清。2、裂解: 加入1×RBC Lysis Buffer，轻柔吹打混匀，裂解1～2min。 3、离心，弃红色上清。本步骤亦可在室温下操作。4、如果发现红细胞裂解不完全，可以重复上述
煮沸后的蛋白加入sds－buffer的储存方法
可以蛋白电泳1×loadingbuffer可以配制出来，但是却很难在实际使用的时候用上比如说蛋白电泳5×loadingbuffer，在使用的时候也就是1个体积的loadingbuffer要加上4个体积的蛋白样品。这样最后的蛋白电泳loadingbuffer就被稀释到1×loadingbuffer如果
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
Restriction－digestion
Restriction digestionRestriction enzyme digestions are performed by incubating double-stranded DNA molecules with an appropriate amount of restriction
常用试剂配制－4
Phytohemaglutinin (PHA)Available as kidney bean lectin. It is typically used as a stock solution of 10-20 g/ml in balanced salt solution. For tissue c
Methods－for－the－Measurement－of－a－Bacterial－Enzyme－Activity－in－Cell－Lysates2
Measurement of ACTase activityNuclear magnetic resonance spect roscopy (NMR)The unique potential of NMR spectroscopy for monitoring simultaneously the
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants主要试剂 Protein
Indirect－ELISA
实验概要This protocol describes a method of indirect ELISA.主要试剂1. Phosphate buffered saline (PBS) tablet: 10 mM phosphate buffer, pH 7.4, 150 mM NaCl
Dephosphorylation－...
实验概要The method provides a protocol for removal of phosphate groups from proteins, before or after blotting. To demonstrate the specificity of an ant
Cell－Staining－for－SenescenceAssociated－betaGalactosidase－－Activity
DescriptionCell Staining for Senescence-Associated â-Galactosidase (SA-â-Gal) Activity Procedure1. Carefully remove the growth media from the cell cul
Detection－of－Glycoproteins－on－Blot
Detection of Glycoproteins on BlotSource: Contributed by Sharad Purohit, Paller's LabReagentsSodium acetate Buffer (200mM, pH 5.5): Prepare a 200
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants. It should
Capture－ELISA
实验概要Capture ELISA (also known as "sandwich" ELISA) is a sensitive assay to quantitate picogram to microgram quantities of substances (such as hormon
Isolation－of－Actin－and－Myosin－Filaments
LEVEL IIIMaterialsRelaxing Solution0.1 M KCl0.001 M MgCl5 mM ATP0.016 M NaHPONaHPOAdjust pH to 7.30.05 M Sodium phosphate buffer, pH 7.00.001 M EDTA (
红细胞裂解液（RBC－Lysis－Buffer,10×）使用说明
红细胞裂解液（RBC Lysis Buffer,10×）简介在生物科研领域，经常需要去除红细胞。去除红细胞的方法有多种，如ACK Lysis Buffer、Tris-氯化铵红细胞裂解液、Gey＇s Lysis Buffer。红细胞裂解液（Red Blood Cell Lysis Buffer
提取质粒用的5种buffer分别有什么作用
提取质粒用的5种buffer分别作用：buffer P1：除去RNA。buffer P2：裂解细胞。buffer P3：沉淀DNA。buffer WA、buffer WB：都是洗涤液。TE：溶解DNA。
关于磷酸缓冲液的简介
磷酸缓冲液（PB，Phosphate Buffer），是生物化学研究中使用最为广泛的一种缓冲液，通常使用的有磷酸钠缓冲液（NaH2PO4&Na2HPO4）和磷酸钾缓冲液（K2HPO4&KH2PO4），由于它们有二级解离，缓冲的pH值范围很广。 主要作用：PB(Phosphate Buffere
流式细胞术检测细胞凋亡
实验概要Provides a rapid and convenient assay for apoptosis. 实验原理Apoptosis is a carefully regulated process of cell death that occurs as a normal part o
Acetylation－(or－Succinylation)－of－Amino－Groups－on－Proteins
Acetylation (or Succinylation) of Amino Groups on ProteinsREFERENCE: Hanock and Benz. 1986. BBA. 860:699-707.PURPOSE: Derivitization of amino groups t
Indirect－ELISA
实验概要 In the indirect ELISA, the enzyme-antibody conjugate uses an antibody against the type of antibody that is used to detect the antigen, kind of
Fixation－and－Embedding－of－Microtubules－for－Electron－Microscopy
(This procedure can also be used for virtually any material that must be pelleted prior to fixation and thin sectioning)Primary fix:2% glutaraldehyde
Radioiodination
Purpose and backgroundsPrinciple of radioiodinationAddition of oxidizing reagents (such as chloramine-T or peroxidase+H202) converts I- to I+ or I3-.
Radioiodination－of－protein
Radioiodination (by Jun Takagi,6/16/2000)Purpose and backgroundsPrinciple of radioiodinationAddition of oxidizing reagents (such as chloramine-T or pe
porcine－brain－tubulin－prep
Although many protocols for tubulin preparation are available, the procedure described below is the simplest and highest yielding preparation I have d
Preparation－of－tubulin
Although many protocols for tubulin preparation are available, the procedure described below is the simplest and highest yielding preparation I have d
troponin蛋白纯化－Protein－purification:－troponins
Overview TROPONINS The calcium-dependent regulatory protein complex located on the thin actin filaments of muscle comprises of TnC (17.8 kDa), TnI (2