Apoptosis:Miniassay
1) Aliquot 5 X 106 total cells in 5-8 ml 2% media.2) Add 30 µl 3H-thymidine.3) Incubate for approximately 16 hours under normal growth conditions.4) Spin down cells and wash 2-3 times with PBS (or serum free media) to remove any unincorporated label.5) Resuspend cells at 5 X 105/ml in serum free media.6) Aliquot approximately 400 µl of cell suspension into each well of a 24 well plate.7) Treat cells (ALWAYS HAVE TIME......阅读全文
Apoptosis:－Miniassay
1) Aliquot 5 X 106 total cells in 5-8 ml 2% media.2) Add 30 µl 3H-thymidine.3) Incubate for approximately 16 hours under normal growth conditions.4) S
Apoptosis－Induction
IntroductionWhen studying induction of apoptosis via a cell surface molecule, it is important to first ascertain surface expression of the molecule of
Chemical－Induction－of－Apoptosis
Chemical Induction of Apoptosis - 1 May 2001p53, p21WAF1, Myc, Bcl-2, Bax, Bcl-x and bak are among the proteins involved in the regulation of apoptosi
Caspase－Cascade－in－Apoptosis
Apoptosis, programmed cell death, is triggered by a variety of stimuli, including cell surface receptors like FAS, mitochondrial response to stress, a
DNA－Fragmentation－Assays－for－Apoptosis
Protocol I: Triton X-100 Lysis BufferIn 96 flat-wells plate, incubate 4x10 6 target cells (40 wells of 105 per well) with desired concentration of eff
Granzyme－A－mediated－Apoptosis－Pathway
One mechanism used by cytotoxic T cells to kill tumor cells and virus-infected cells is the release of perforin and granzyme proteins. Perforin protei
FACS－Procedures－for－Apoptosis－Detection
Materials:Hoechst 33258 (Sigma B-2883).stock: 10 mg/ml in dH20 (40)working dilution: 500µg/ml (50µl stock + 950µl PBS).7-Amino-actinomycin (Sigma A-94
Free－Radical－Induced－Apoptosis
Oxidative stress is one factor that can trigger programmed cell death. Activated neutrophils responding to inflammatory stimulation produce reactive o
HIV－Induced－T－Cell－Apoptosis
HIV infection is associated with immunosuppression caused by a dramatic reduction in the helper T cell population. The loss of helper T cells may be c
Apoptosis－TUNEL－Assay－(frozen－sections)
Protocol for Frozen Sections:Warm 150ml 4% Paraformaldehyde/1x PBS to RT. Fix slides in it, 20 min., RT.1x PBS rinse, 2 times.1x PBS, 30 min., RT. Beg
Apoptosis－TUNEL－assay－(Paraffin－Sections)
Protocol for Paraffin Sections:Dewax paraffin sections:Incubate slides, 55°C, 30 min.Xylenes, 2 times, 2 min. each100% EtOH, 2 times, 2 min. each95% E
Guide－to－Cell－Proliferation－and－Apoptosis－Methods
Chapter 1: Cell Death - Apoptosis and Necrosis1.1Introduction21.1.1Terminology of cell death21.1.2Differences between necrosis and apoptosis31.1.3Apop
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement
IntroductionApoptosis is a normal physiological phenomenon put forward by Kerr [1]. It plays an important role in embryonic development, maintenance o
Opposing－roles－of－AIF－in－Apoptosis－and－Cell－Survival
Programmed cell death is induced by many different factors and involves numerous signaling pathways, some dependent on caspase proteases and others th
A－rapid,－quantitative－and－inexpensive－method－for－detecting－apoptosis
ASTRACTWe describe a rapid and quantitative flow cytometric method for determining the apoptotic or anti-apoptotic potential of a gene in various cell
细胞凋亡（apoptosis）的检测3
【结果判断】细胞出现程序化细胞死亡时，在琼脂糖凝胶电泳带上呈现有一定间隔的梯状条带，而细胞坏死时则呈模糊的片状条带（无间隔），阴性对照者仅在近电泳点样处出现基因组条带。（3）流式细胞仪观察－PI染色法【原理及意义】细胞凋亡时，流式细胞检测可呈现亚二倍体核型峰的特征。此外，根据细胞光散射特点，应用碘化
细胞凋亡（apoptosis）的检测2
【材料】１试剂：吖啶橙贮存液（10mg吖啶橙溶解于100mLPBS中，过滤，4℃避光保存），0.01mol/L、Ph6.8PBS。２器材：吸管，试管，玻片，荧光显微镜。【方法】１制备待检活细胞悬液，浓度约为1×10７/mL。２取95uL的细胞悬液，加5uL的吖啶橙贮存液混匀。３吸一滴混合液
细胞凋亡（apoptosis）的检测1
细胞凋亡（apoptosis）是一种由基因控制的细胞自主性死亡方式。其发生诱因及形态学特征均有别于坏死，与人类免疫系统的发生发展、神经组织发育、器官的形成、肿瘤的发生发展等诸多生物学现象之间有着密切联系，并且是免疫应答过程中免疫细胞的杀伤机制之一。细胞凋亡的检测技术已成为免疫学检测的一个重要内容。凋
Role－of－nicotinic－acetylcholine－receptors－in－the－regulation－of－apoptosis
Nicotinic acetylcholine receptors are essential for neuromuscular signaling and are also expressed in non-neuronal tissues, where their function is le
PTEN－dependent－cell－cycle－arrest－and－apoptosis
PTEN is a tumor suppressor gene. Recombinant PTEN is capable of dephosphorylating phosphatidylinositol 3,4,5-triphosphate[PI(3,4,5)P3], the product of
Apoptosis:－A－Laboratory－Manual－of－Experimental－Methods－Andrea－Cossarizza
THE CELL 1. Morphological aspects of apoptosis Walter Malorni, Stefano Fais & Carla Fiorentini 2. Cell cycle Miriam Capri & Daniela BarbieriTHE NUCLEU
Detection－Of－Cell－Viability－And/Or－Apoptosis－By－Flow－Cytometry－(FACS)
Viable cells are cells that when allowed to continue beyond the timepoint of examination will stay alive. Besides live and healthy cells, cells in ear
In－Situ－Cell－Death－(Apoptosis)－Detection－by－TUNEL－labeling
Protocol for Frozen Sections:Warm 150ml 4% Paraformaldehyde/1x PBS to RT. Fix slides in it, 20 min., RT.1x PBS rinse, 2 times.1x PBS, 30 min., RT. Beg
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement2
TroubleshootingCritical Steps(1) Don’t trypsinize cells for too long when collecting them.(2) Rotation speed should be no more than 1500 rpm during ce
细胞凋亡（Apoptosis）检测技术与方法
细胞凋亡（Apoptosis），又称细胞程序性死亡（PCD： Programmed Cell Death），是指细胞在一定的生理或病理条件下，遵循自身的程序，自己结束其生命的过程。它是一个主动的，高度有序的，基因控制的，一系列酶参与的过程。细胞凋亡概念提出至今还不到30年的时间，但由于它在保证多
ASENSITIVE－METHOD－FOR－DETECTION－OF－APOPTOSIS－BY－SINGLE－LASER－FLOW－CYTOMETRY
MATERIALS:1. 1 X PBS (PBSAz, 1 X PBS, e.g., Irvine Scientific, CA, containing 2% newborn calf serum and 0.1% sodium azide)2. 7-Amino-actinomycin D (7-
A－rapid,－quantitative－and－inexpensive－method－for－detecting－apoptosis2
Figure 1.Determination of apoptosis in transiently transfected murine [beta] tumor cells. (A) The number of apoptotic [beta]HC 13T tumor cells (% apop
Neuropeptides－VIP－and－PACAP－inhibit－the－apoptosis－of－activated－T－cells
Vasoactive intestinal peptide (VIP) and the structurally related pituitary adenylate cyclase-activating polypeptide (PACAP), two neuropeptides present
TSP1－Induced－Apoptosis－in－Microvascular－Endothelial－Cell
As tissues grow they require angiogenesis to occur if they are to be supplied with blood vessels and survive. Factors that inhibit angiogenesis might
Role－of－MEF2D－in－Tcell－Apoptosis
Mef2 (Myocyte enhancer factor 2) transcription factors play a role in T-Cell Calcium Induced Apoptosis. Several factors regulate Mef2 transcription fa