Purificationofhumanmononuclearcellsandneutrophils
PurposeMaterials10ml 6% dextran + 7ml citrate/citric acidDextran: T500 --> 6g+100ml PBSCitrate solution: 25g Na Citrate + 8g citric acid + 500 ml PBS43 ml blood12 ml RT Histopaque 107718 ml cold H2O2 ml 10x PBSM199 for HUVECs: 1L powder pocket M199 + 35g NaHCO3 + 25mM Hepes + 5 mM glutamine + 50 ug/ml Gentamycin.1M Hepes: 119.1 g Hepes + 500 ml dH2O.Media A: 1X HBSS (10X:50ml) + 10 mM Hepes (1M: 5 ml) + 5 mM EDTA ......阅读全文
Isolation－of－human－multipotent－mesenchymal－stem－cells－from－second
Isolation of human multipotent mesenchymal stem cells from second‐trimester amniotic fluid Culture of MSC from amniotic fluid1. Twenty amniotic flui
Isolation－and－longterm－cultivation－of－Human－Corneal－Endothelial－Cells
Coating of the Culture Dish SurfacesCulture dishes were incubated with a film of a 1:1 mixture of laminin (10 μg/ml and chondroitin sulfate (10 μg
Isolation－and－Culture－of－Multipotent－Stem－Cells－from－Human－Bone－Morrow
Bone marrow contains three types of stem cells:1. Hematopoietic stem cells give rise to the three classes of blood cells that are found in the circu
Ex－Vivo－Human－Primary－Mesenchymal－Stem－Cells－(MSCs)－Culture
Human bone marrow contains mesenchymal progenitors (mesenchymal stem cells, MSCs). MSCs produce adventitial cells in the human bone marrow microenvir
Culture－of－human－renal－proximal－tubule－cells－and－renal－cortical－fibroblasts
1. The method for primary culture of human renal proximal tubule cells (PTC) and renal cortical fibroblasts (CF) is described in detail.2. Renal cor
Preparation－of－Mouse－Neutrophils
实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline solution [HBSS; Invitrogen, Grand Isla
Preparation－of－Mouse－Neutrophils
实验概要Preparation of Mouse Neutrophils实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline so
Procedure－for－Culturing－BG01V－Human－Embryonic－Stem－Cells
IntroductionHuman embryonic stem (hES) cells are pluripotent stem cells derived from pre-implantation embryos that can be maintained and expanded in a
Examination－of－a－Mammalian－Blood－Smear
Examination of a Mammalian Blood SmearThe distribution and appearance of the formed elements of blood can tell a great deal about the condition of the
3－Color－Staining:－AlphaCatenin－in－HeLa－Human－Cervical－Cancer－Cells
实验概要Alpha-catenin in HeLa human cervical cancer cells was labeled using mouse anti-α-catenin and visualized with Alexa Fluor® 488 goat anti-mouse Ig
Purification－of－Lin,－ckit+,－Sca1+－bone－marrow－cells－for－Culture
MATERIALSMedia:Heat inactivated FBS (56°C x 30 min)PBS + 2% heat-inactivated FBSIMDM + 20% heat-inactivated FBSViral transduction: Iscove's + 20%
Phycoerythrin－conjugation－protocol
Phycoerythrin conjugation protocolDavid's method modified from references (2) and (3). I used this method to conjugate a mouse IgG2a monoclonal an
Culturing－BG01V－Human－Embryonic－Stem－Cells－with－Mouse－Embryonic－Fibroblast
If culturing in the absence of a feeder cell layer is desired, human embryonic stem (hES) cells can be maintained using Mouse or Human-Conditioned Med
ex－vivo－expanded－endothelial－progenitor－cells
Cell Culture. 1. Total hPBMCs were isolated from blood of human volunteers by density gradient centrifugation. 2. Cells were plated on culture dishes
Hematopoietic－Stem－Cell－Targeting－with－Surface1
Hematopoietic Stem Cell Targeting with Surface-Engineered Lentiviral VectorsEls Verhoeyen and François-Loïc CossetAdapted from Gene Transfer: Delivery
碳水化合物分析
Carbohydrate Assay (Hancock Laboratory) (Accessible only by IE)This protocol is used to determine the relative amounts of LPS CHO present in a given s
DNA微序列技术
· Protocols for Making Drosophila Arrays (Stanford U.)Detailed protocol for making arrays including PCR Amplification of cDNAs for Printing,
fMLP－induced－chemokine－gene－expression－in－HMC1－cells
Neutrophils respond to bacterial infection by releasing reactive oxygen species that kill bacteria and by expressing chemokines that attract other imm
Antibody－Purification
This protocol includes an ammonium sulfate cut, affigel blue chromatography and affinity chromatography.1. Solutions(1) Affigel Blue Prewash0.1 M acet
中山大学最新《Blood》文章解析细胞因子新发现
郑利民 教授 1984年毕业于上海医科大学后在附属华山医院任临床医生，94年获荷兰莱顿（Leiden）大学细胞生物和免疫学博士学位，1999年获瑞典国家研究院人才基金（助理教授）资助而在瑞典林雪平大学建立了独立的实验室。2002年被聘为中山大学教授，博士导师。现任中山大学生命科学院生物化学系主
PriCells:－Isolation－of－endothelial－progenitor－cells－(EPCs)
PriCells: Isolation of endothelial progenitor cells (EPCs) 1. Twenty-four mL venous blood was collected at each time point into Vacutainer CPT Mono
Neutrophil－and－Its－Surface－Molecules
Neutrophils are important phagocytotic leukocytes (white blood cells) that internalize and destroy infectious bacteria by a respiratory burst of react
Isolation－of－liver－lymphocyte
Isolation of liver lymphocyte Several lymphocyte subpopulations reside in the normal adult human liver. Liver lymphocytes mainly include a large n
Placental－trophoblast－and－chorionic－cell－cultures
1. Placental trophoblast and chorionic trophoblast cells were prepared using a modification of the method. 2. Term human placentae and chorion tissue
Human－B－cell－isolation－and－culture
实验概要This protocol provides a general protocol for human B cell isolation and culture.实验步骤B cell isolation1. Donor blood was obtained with informed con
Protein－purification;－actin
Protein purification; actin Overview ACTINThe most abundant muscle and non-muscle cytoskeletal protein. MW 42 kDa, 374/375 amino acids; various
Purification－of－mAb－(IgG)
1. Materials(1) Antibody 7E3, 2L sup grown in flasks, frozen and thawed overnight.(2) BioRad Affi-Gel Protein A MAPS II Buffers cat. #1530-6160 ($161.
Protein－A－Purification－of－Antibody
1. Reagents(1) Affi-gel Protein-A Agarose (BioRad #153-6153)(2) MAPS II Binding Buffer (BioRad # 153-6161)(3) 0.314 g/ml diH2O(4) MAPS II Elution Buff
Purification－of－Demethylated－Sphingomyelin
I. Lower, chloroform phase:1) Dry on rotovapor system with house vacuum lines. It is not necessary to dry sample completely, but sufficiently to yield
The－UnderAgarose－Migration－Assay
overviewThe Under-Agarose assay is a useful method for observing the response of a cell population to one or more chemoattractant sources. The behavio