FreezingandThawingofMammalianCellLines
For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture.FreezingPreparationCells are to be frozen in liquid nitrogen, so make sure your canisters are relatively full of nitrogen and you have room. Cells should be healthy (>90% viability) and growing in log phase. You will also require sterile 1 mL cryo-vials; they have a scr......阅读全文
哺乳动物RNAi技术全面实验方法
WHENVIRUSESINFECTEUKARYOTICCELLSdomly integrate into host genomes, double-stranded RNA (dsRNA) is frequently produced from the invading genes, either
Preserving-yeast-cultures
Short term storageYeast cultures are stable for 1-2 weeks when refrigerated. Petri dishes should be sealed or in plastic bags.Medium term storageYeast
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
The-OP9DL1-System:-Generation-of-TLymphocytes-from-Embryonic4
TROUBLESHOOTINGProblem: The OP9 cells are more than 80%-90% confluent.Solution: It is important when creating working stocks of OP9 cells for freezing
SDS-Whole-Cell-Extracts
-Use sterile technique and sterile solutions in steps 1 to 3.-1. Using a saturated starter culture, inoculate 25 to 30 ml of appropriate media in a 12
Purification-of-Genomic-DNA-Using-PureLink™-Silica-Columns
实验概要The PureLink™ Genomic DNA Purification Kit allows rapid and efficient purification of genomic DNA. The kit is designed to efficiently isolate ge
线粒体荧光探针大全:TMRM,Mitotracker,JC1(5)
Monoclonal Antibodies Specific for Proteins in the Oxidative Phosphorylation SystemOxidative phosphorylation (OxPhos) activity occurs in the mitochond
Yeast-Genomic-DNA-Prep
Grow 10ml YPD cultures o/n. Figure out cell density; inoculate 30 ml YPD and grow o/n so that cell density is approximately 2 x 108cells/ml the next m
Master-Cell-Bank
PurposeTo describe the preparation of a Master Cell BankSafetySee SP 09-001 for lab safety considerations for the cell culture lab.EquipmentLaminar Fl
alamarBlue®-Cell-Viability-Assay-Protocol
实验概要Cell health can be monitored by numerous methods. Plasma membrane integrity, DNA synthesis, DNA content, enzyme activity, presence of ATP, and c
PBMC细胞的精确计数和活性分析(三)
五、使用仪器发表文章AuthorDateTitleJournalCell TypeCellometer / ApplicationsMahato, Ram INovember 2013Synthesis and Characterization of an Anti-Apoptotic Immu
报告基因检测的精明之选—LipoD293DNA转染试剂
使用lipoD293转染试剂来转染哺乳动物细胞(比如Hela 、PC3),以期检测荧光酶报告基因,同其他转染试剂相比较,您会得到更多的荧光酶读数,更少的细胞死亡率。使用unsupplemented RPM11640/DMEM替代Opti-MEM培养基来稀释lipoD293及DNA(luciferas
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
CO2恒温摇床解决人胚肾-293-(HEK293)-细胞结团问题(一)
人胚肾 293 (HEK293) 细胞在重组蛋白表达中是最常见的宿主细胞。 这类细胞能够表达大量的膜蛋白,如 G 蛋白偶联受体 (GPCR) ,是无法在最常见的生物制药生产宿主,如:中国仓鼠卵巢 (CHO) 细胞中作表达。 HEK293 虽然是蛋白表达的极好宿主,然而 HEK293 细胞
Working-Cell-Bank
1) Keep prepared solutions on ice.2) Determine total cell count of cells to be frozen. (e.g. 1 X 108 )3) Determine number of vials to be frozen. (e.g.
The-OP9DL1-System:-Generation-of-TLymphocytes-from-Embryonic1
The OP9-DL1 System: Generation of T-Lymphocytes from Embryonic or Hematopoietic Stem Cells In VitroRoxanne Holmes and Juan Carlos Zúñiga-Pflücker1Sunn
InVitro-Adipocytes-Differentiation
IntroductionObesity is a significant clinical problem that contributes to life-threatening diseases such as diabetes and atherosclerosis. With an incr
Growing-cells
No two cell lines behave exactly the same, so you must learn the peculiarities, or personality, of each of the cell lines with which you work. Irrespe
Sauer:Lysing-E.-coli-with-Lysozymes
Getting The Most Out Of Your BugsNative lysis is a staple protocol in practically every biochemistry lab, yet there is significant variability in the
Method:-Logging-in-Specimens-and-Record-Keeping
Method: Logging in Specimens and Record KeepingJune 10, 1990Rosalie VeilePurpose:To keep a written and computerized record of all cell lines, the date
Midiprep-preparation-of-Plasmid-DNA
实验概要The PureLink™ HiPure Plasmid DNA Midiprep Kit allows purification of 100–350 μg of high-quality plasmid DNA from 15–25 mL overnight E. coli cul
Cryopreserving-Neural-Stem-Cells
实验概要There are numerous protocols available for cryopreserving neural stem cells (NSCs) derived from human embryonic stem cells; the primary objectiv
Optimized-Method-for-the-Preparation-of-Rodent-Testicular-Cells1
Homogeneity of cell populations is a prerequisite for the analysis of biochemical and molecular events during male gamete differentiation. Given the c
PC12-Cell-Culture-and-Fusion
Cell CultureMaterials1. Falcon Primaria culture dishes.2. Culture medium: DME (or F12K) with glutamine, supplemented with 7% heat-deactivated horse se
Cell-Viability-Assay
Dye exclusiona cell suspension is mixed with trypan blue and examined by low-power microscopyMaterialscellsPBSM3hemocytometer0.4 % trypan blue in PBSm
CellTrace™-CFSE-Cell-Proliferation-Kit
实验概要The CellTrace™ CFSE Cell Proliferation Kit provides a versatile and well-retained cell-tracing reagent in a convenient and easy-to-use form. The
siRNA用户手册
The siRNA user guide (revised May 6, 2004) Selection of siRNA duplexes from the target mRNA sequenceUsing Drosophila melanogaster lysates (Tuschl et
SiRNA用户指南
Selection of siRNA duplexes from the target mRNA sequenceUsing Drosophila melanogaster lysates (Tuschl et al. 1999), we have systematically analyzed t
MicroRNA-Expression-Profiling-by-Bead-Array-1
MicroRNA Expression Profiling by Bead Array Technology in Human Tumor Cell Lines Treated with Interferon-Alpha-2aMicroRNAs are positive and negative r
TEM-Specimen-Preparation:Preparative-Techniques-for-the-TEM
For routine transmission electron microscopy (TEM), it is generally accepted that specimens should be thin, dry and contain molecules which diffract e