NeedleAssayforChemotaxis
Devreotes Lab, John Hopkins Medical Institutions http://www.hopkinsmedicine.org/cellbio/devreotes/needle.htmEquipment and chemicalsZeiss inverted microscope, eppendolf microinjector, eppendolf micromanipulator, (in Dr. Murphy’s lab)Eppendolf femtotips, 100mM cAMP in PM,DB buffer, PM Buffer.Preparing Cellsa. Wash cells with DB and resuspend them to 2 X 107 / mlb. Shake cells at 120 rpm for 1 hourc. Pulse cel......阅读全文
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
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
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
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
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.
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
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
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
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-
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
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
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
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
Nucleotide－Binding/Hydrolysis－Assay
MaterialsNucleotide mixMotor (50 - 100 µM; purity > 95%)0.5 M Tris-OAc, pH 7.510 mM EGTA10 mM MgCl2DDWSephadex G-50 Medium column (0.8 cm in x 20 cm)C
Angiotensin－Protein－Kinase－Assay
James Hardwick's angiotensin assay protocolThis specific procedure was developed to assay the activity of the Lck kinase expressed from a retrovir
Cr－Release－Cytotoxicity－assay
DescriptionCytotoxic activities of mNK cells are examined in 51Cr release assay from target cells ProcedureEffector cells (mNK cells) are seeded into
Polyphenoloxidase－(catechol－oxidases)－assay
Browning of the cut surface of some fruits and vegetables is due the presence of a group of enzymes called polyphenoloxidases. These enzymes are relea
Biorad－Protein－Assay:－Bradford
Biorad Protein Assay: BradfordStandards: 1 mg/ml BSA stock- dilute 1:10 to get 0.1 mg/ml BSAAdd To get H-2O20 µl 2 µg/ml 780 µl40 µl 4 µg/ml 760 µl60
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
Chorioallantoic－Membrane－(CAM)－Assay
8 eggs per day, day 7- day 13 cut CAM, wash in precooled PBS,in 10 ml WASH 1 (PBS, 5 mM EDTA, COMPLETE) on icecut in pieces in petri dish on icecentri
Xenograft－Tumor－Assay－Protocol
1) Determine the number of cells for injection (ie 5´106 ) to determine the number of plates thatwill require trypsinizing (usually a 100% confluent p
Gel－Shift－Assay－Systems
ProtocolsDownloadprotocol183kbpdf?Abstract for Gel Shift Assay SystemsThe gel shift, or electrophoretic mobility shift, assay provides a simple and ra
AlamarBlue®－Cell－Viability－Assay
实验概要Assess cell viability. 实验原理Cell health can be monitored by numerous methods. Plasma membrane integrity, DNA synthesis, DNA content, enzyme activ
Phosphate－Assay－by－Suprya－Jaydev
ReagentsAshing buffer:10 g Mg(NO3)2 100 ml EtOH1.5 N HCl stock:119.7 ml concentrated HCl (11.6M @ 36% by weight)880.3 ml H2O1 N Sulfuric acid stock:28
Protocol－for－Aortic－Ring－Assay
ProceduresCover a 48-well plate with Matrigel (100μl/well) of and incubate for 30 min at 37°C, 5% CO2.Sacrifice the1-2 month old mice/rats (WT/mutant
Hanging－drop－aggregation－assay
DescriptionThis assay is used for the aggregation property of cancer cells. It is a very critical parameter for measurement of cell metastasis. Factor
Cell－Clonogenic－Survival－Assay
DescriptionAllows one to test the capability of adherent cells to survive and replicate following insult with chemicals or radiation. Procedure1. Grow
Bradford－Protein－Concentration－Assay
Bradford Protein Concentration Assayversion 01/07/2001Abbreviations:mcg = microgramsmcL = microlitersBSA = bovine serum albuminO.D. = optical densityd