Isolated cells grown in optimal tissue culture conditions will most likely undergo apoptosis once placed in a foreign environment.
To determine whether the cell undergoes an intrinsic or extrinsic apoptotic pathway by observing the amount of caspase 8 activated.
Human adherent cells are used for the tissue culture. They are washed carefully and loose cells are isolated. Adherent cells are gently spun and labelled with FAM green fluorescence from the FLICATM kit before trypsinization. Cells are incubated with the FLICA at 37oC for roughly 60 minutes before washing. After the addition of apoptosis wash buffer, cells are placed on ice and protected from light. Observations are made by the analysis of cells with a flow cytometer.
Click ‘Protocol’ tab in the following link for more detailed instructions.
Suppliers: Immunochemistry Technologies
Resource: Green FLICA™ Caspase 8 Assay Kit
If there is a high fluorescence reading observed, that means that there is a high amount of caspase 8 present and therefore, leads to an extrinsic apoptotic pathway. However, if there is a low fluorescence reading observed, there is less caspase 8 and hence, there is no extrinsic pathway activated.
-The Introduction was nice and brief and gave a good overview on phagocytosis.
-The links fitted in nicely, helped outsource information.
-AS I am not a scientist probably better off explaining some of the short hand names.
-Receptors could be expanded upon.
-Nice comparison with the 2 types of models introduced. I think this could be enhanced by a picture of each model (not just one of them), as some people are visual learners and can use the pictures to help understand.
-The mechanism had good information, although some of the sentences didn’t flow.
-In the disease section, the references should be placed after key information and not after.
-I like the fact they had a few diseases to talk about (which gives me ideas for my project), maybe just need to go a little bit more in depth of certain activities and causes.
-With the referencing, I think a lot could be improved and this would help the information and flow of the project be formatted much better.
-Also one reference section at the end rather than 2.
-Some references were used twice, which is fine but doubled up in the reference list for some reason..
-I think overall, this project has good information, it just needs some adjusting in the formatting and referencing departments.
-I think images would be useful in describing pathway the mechanisms involved.
-The names should be stated before the names are written in shorthand, so that the reader can know the full name. The information on them is good though.
-More information needed on the structure and function of the Rans mentioned.
-More headings needed.
-Expand the diseases, go into a bit more detail.
-With the referencing, need to fix up the list a little bit. There are some which appear twice on the list, so a quick edit will do the job.
-I think, overall this project has good information, which needs to be expanded upon and the use of images would really enhance the page.
-The introduction lacked information. The mechanisms are basically the same, so not sure if there is a need to individualise the various transports.
-The transports were well written, just an image/diagram would help visualise the mechanisms involved.
-The table for the proteins were an added benefit for the reader.
-Before the use of short hand names, the name should be introduced so the reader is aware of the actual name of the product.
-More information should be used to explain the headings involved.
-Images need to be spread amongst the project page and not centralised at the bottom. Therefore some of the images actually correlate to above headings but are at the bottom of the page where there is no correlation.
-In the referencing there were a few mistakes with putting in the right format.
-I think overall, this project needs to add a fair amount of information, better format with more headings and subheadings. I think they are on the right track for a good project.
The Anti-TOMM20 antibody, Product code ab56783, is a monoclonal antibody raised in mouse. The antibody reacts with rat and human. The applications that can be used with this antibody are; western blotting, immunohistochemistry and immunofluorescence.
Zhang B et al. SIRT3 overexpression antagonizes high glucose accelerated cellular senescence in human diploid fibroblasts via the SIRT3-FOXO1 signaling pathway. Age (Dordr) 35:2237-53 (2013). ICC/IF ; Human .<pubmed>23494737</pubmed> Ramonet D et al. Optic atrophy 1 mediates mitochondria remodeling and dopaminergic neurodegeneration linked to complex I deficiency. Cell Death Differ : (2012).<pubmed>22858546</pubmed> Steketee MB et al. Mitochondrial dynamics regulate growth cone motility, guidance, and neurite growth rate in perinatal retinal ganglion cells in vitro. Invest Ophthalmol Vis Sci 53:7402-11 (2012). PubMed: 23049086 Verfaillie T et al. PERK is required at the ER-mitochondrial contact sites to convey apoptosis after ROS-based ER stress. Cell Death Differ 19:1880-91 (2012).<pubmed>22705852</pubmed> Munday DC et al. Quantitative proteomic analysis of A549 cells infected with human respiratory syncytial virus. Mol Cell Proteomics 9:2438-59 (2010). ICC/IF ; Human . PubMed: 20647383 Darna M et al. Time of day-dependent sorting of the vesicular glutamate transporter to the plasma membrane. J Biol Chem 284:4300-7 (2009). WB ; Rat .<pubmed>19103593</pubmed> Whittaker R et al. Kinetics of the translocation and phosphorylation of alphaB-crystallin in mouse heart mitochondria during ex vivo ischemia. Am J Physiol Heart Circ Physiol 296:H1633-42 (2009).
This article talks about how a mitochondrial dysfunction is a common characteristic in parkinson disease. The disease gene PINK1, results in deterioration of the functions of mitochondria. PINK1 is also found in the cytoplasm and when overexpressed enhanced cell motility. 
Mutations in PTEN-induced putative kinase 1 (PINK1) are a cause of autosomal recessive familial Parkinson's disease.  Attempts in studying the PINK1 pathway have been controversial because of its location in the cell and within the mitochondria. This study shows that this protein has a topology that the kinase domain faces the cytoplasm and the N-terminal tail is in the mitochondria. The results within this study rectify the location of PINK1 in mitochondria and may help in realising the normal and physiological function and normal physiological function and potential pathogenic role in Parkinsons disease. 
PINK1 is a serine/threonine kinase in the outer membrane of mitochondria.  It is referred to as the gene responsible of Parkinson's disease. In the cytosol is where PINK1's precursor is made and then transported into the mitochondria through the TOM complex. It discusses the fact that the import receptor Tom70 is essential for PINK1 import. It also states that the study observed that PINK1 has predicted mitochondrial targeting signal. Thus, the research results suggest that PINK1 is transported to the mitochondria by a pathway that is independent of the TOM core complex but crucially depends on the import receptor Tom70. 
PINK1 mutations are connected to autosomal recessive parkinsonism. PINK1 influences the shape and the function of the mitochondria. this paper describes a mechanism linking the dysfunction of the mitochondria and the changing of the mitochondrial shape that is related to PINK1. 
2.In vivo, confocal microscopy is being used more and more to study corneal abnormalities. Using this, we can find drug deposits in systemic disease. This article talks about a woman diagnosed with fibromyalgia on systemic choloquine who is going for an ophthalmic examination. 
This is about prokaryotes.
this is great