ActinCaptureAssay
David AmbergDialyze purified GST fusion proteins and actin into PBS + 1mM MgCl2 .Mix 5ug actin into 50ul total volume binding buffer.Mix 5ug GST-fusion protein into total volume 50ul binding buffer.Combine actin + GST-fusion protein and incubate for 1 hr. x 4¡C.Add 20ul 50% glutathione agarose equilibrated into binding buffer. Incubate 15 min x 4¡C with frequent mixing.Wash beads 4 times with binding buffer, spin dow......阅读全文
Assay－for－the－Micrococcal－Nuclease
Method: Essentially that described by Heins et al. (1966) based upon the release of acid soluble oligonucleotides following nuclease digestion of DNA.
Glucosamine－Rapid－Assay
Glucosamine Rapid AssayMRTHOD:Place sample (containing 0.5 - 10 µg GlcN) in a Pyrex screw capped tube.Add HCl to a final concentration of 2N and a fin
LOWRY－PROTEIN－ASSAY
The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al., J. Biol. Chem. 193: 265-
Bradford－protein－assay
Bradford protein assayConsiderations for useThe Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is fairly
Tube－formation－assay
DescriptionThis is a fast and easy assay to test the angiogenic/anti-angiogenic properties of molecules. As compared to other angiogenesis assays, suc
Noble－Agar－Assay
DescriptionCancer cells do not show anchorage and contact inhibition of growth. To assess the anchorage and contact independent growth of cells, noble
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
DNA－methyltransferase－Assay
Methylated CpG island Amplification Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polymerase
Migration－Assay－Protocol
Materials to be prepared beforehand:1) FBS free medium2) 10% FBS medium3) Cell migration filter insert ( Transwell®, 12mm Diameter, 12 μm Pore Size.)P
Adhesion－Assay－Protocol
Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Lamini
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 an
Pheromone－Halo－Assay
-Use sterile technique and sterile solutions throughout this method.-1. Grow a starter culture at 30 C with shaking (250 rpm) until it reaches saturat
HISTONE－KINASE－ASSAY
PROTOCOLTo 1.5 mL eppendorf tubes add:200 µg of protein extract (see Western blot protocol for protein sample preps)q.s. to 300 µL with RIPA (with pro
Protein－Assay－(Spectrophotometer)
Protein Assay (Spectrophotometer)Use BSA (bovine serum albumin) 1mg/ml stock solution (1ml Eppendorf tubes) for standard curve.Place 0, 2, 5, 10, 15,
Cell－Viability－Assay
Dye exclusiona cell suspension is mixed with trypan blue and examined by low-power microscopyMaterialscellsPBSM3hemocytometer0.4 % trypan blue in PBSm
cell－proliferation－assay
cell proliferation assaybefore start:thaw cells from liquid nitrogen, grow in 75cc flask (T75) in Fischer's medium MM (maintenance medium) until c
ELISA－Inhibition－Assay
ELISA Inhibition AssaySensitize a 96-well microtiter plate with purified antigen.Prepare a solution of the purified antigen of interest in phosphate b
Wound－healing－assay
The wound healing assay allows the researcher to study cell migration and cell interactions. In some cases also single cell migration can be analyzed.
Leaf－GUS－Assay
实验概要a protocol for Leaf GUS Assay This protocol is for small samples (usually single leaf from 21DAI plants), scale up for larger samplesAs there are
In－vitro－Sphingomyelinase－Assay
Reagents:Lysis buffer25 mM Tris-HCl, pH 7.45 mM EDTA1 mM ATP20 µg/ml CLAP1 mM PMSFBuffer A10 mM MgCl20.2 M Tris-HCl, pH 7.40.2 % Triton X-100Buffer B0
Glycolipid－Binding－Assay
Glycolipid Binding AssaySource: Contributed by Pingsunjim, Paller’s LabAbstract: This protocol can be used for the detection of glycolipids binding to
Soft－Agar－Assay
Soft Agar AssayMake 0.6% media-agar mix for the bottom layer. To make 0.6% agar mix the following components (this makes 200 ml):2X DME 100 mlI
Leaf－GUS－Assay
一、实验试剂 GUS Buffer (500 ml) 2.0478 g Na2HPO4 1.2688 g NaH2PO4 (=50 mM NaPi pH7.0) 10 ml 0.5 M EDTA (=10 mM) 0.5 g Triton X-100 0.5 g N-L
Crystal－Violet－Assay
This is a simple assay useful for obtaining quantitative information about the relative density of cells adhering to multi-well cluster dishes. The dy
Needle－Assay－for－Chemotaxis
Devreotes Lab, John Hopkins Medical Institutions http://www.hopkinsmedicine.org/cellbio/devreotes/needle.htmEquipment and chemicalsZeiss inverted micr
ELISPOT－(Enzymelinked－ImmunoSPOT)－实验方法步骤1
IntroductionEnzyme-linked immunosorbent spot (ELISPOT) assays were originally developed to enumerate B cells secreting antigen-specific antibodies, bu
小鼠βactin基因的克隆表达（1）
动物组织RNA的提取实验目的：掌握由动物组织中提取总RNA的方法实验原理：Trizol试剂是一个包含酚、异硫氰酸胍和SDS的单相酸性溶液，其在裂解细胞的同时抑制RNase的活性，随后加入氯仿，酚会大量的溶解在氯仿中。由于DNA和RNA在酸性酚中的溶解性不同，造成DNA分布在下层的氯仿酚溶液中，RNA
小鼠βactin基因的克隆表达（3）
（2）质粒的提取及酶切鉴定分析 质粒的提取按照质粒提取试剂盒的操作步骤进行，然后进行双酶切鉴定。 管号 ① ② 质粒DNA
小鼠βactin基因的克隆表达（2）
实验步骤： （1）目的基因与表达载体的连接反应： ①表达载体和目的片段的酶切 管号 ① ② pGEX 4T-1
如何挑选合适的βActin内参抗体？
如何选择合适的β-Actin抗体？ 现在市场上对于内参抗体选择是比较多，但是也需要遵循一定的原则，那怎么去找到适合自己实验的β-Actin抗体呢？首先，我们需要考虑目的蛋白的大小，我们推荐内参要与检测的目的蛋白的分子量最好相差5KD以上，因此你需要根据你目的检测蛋白的分子量来选择合适的内参，由于β-