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Questions related to Cellular Biology Techniques
Can anyone tell me the difference between the isolation of enzymes from mitcochondria and peroxysomes?
05 March 2015 2,890 3 View
I have done this Cips experiment twice using classic 7 compounds in paper,only showing morphology change of MEF and cloning,no Oct4 activation,no Nanog activation,only showing the reactivation of...
03 March 2015 1,137 0 View
I am trying to carry out experiments which will prove the interaction between two different proteins. One protein is present in the cytoplasm and is thought to be secreted into the environment,...
03 March 2015 3,993 8 View
I'm trying to interface between two phases of a project - the first set of transcriptomic measurements were collected from cells plated on dishes that were not pre-treated. The next phase involves...
03 March 2015 6,466 8 View
I am doing single cell isolation from tumor mass (using collagenase D & DNAse I) for FACS analysis. I also have 1 step for MACS cell separation after collecting single cell suspension....
03 March 2015 5,809 7 View
Dear friends I have cultured hella cells and the cell mortality and viability have been checked under treatment with special drug. So, I have seen that the cell mortality and viability decreased...
03 March 2015 781 8 View
I want to estimate SCC levels in colostrum (1st 2 days) of cow. I am facing difficulty due to thick consistency and presence of fat globules in colostrum. does anyone has experience of estimating...
03 March 2015 4,656 17 View
The hormone binding reaction was initiated by addition of fluorescent tagged natural ligand to each in 6 well plate (4 to 5 lac cells/ per well 6 well plate). Incubations were performed in...
03 March 2015 7,131 4 View
I want to reuse the polycarbonate tubes for isolating exosomes. However i can see some sucrose stuck to the bottom of the ultracentrifuge tube from the previous isolation which is not getting...
03 March 2015 1,417 7 View
what is the optimum time to stimulate PBMCs. i have gone through various manuscripts and they have mentioned various time frames ranging from 2-48 hours. i also came across many manuscripts which...
03 March 2015 5,472 7 View
I'm been trying to measure mitochondrial respiration of the cell line H9c2. I think i been having problem with the permeabilization. I wish to do de analisis using a Clark electrode. Thank...
03 March 2015 4,881 6 View
i'm studying apoptosis and cell cycle arrest induced by a drug, the problem is with some concentrations, after centrifugation at 300g/10min, in the cytometer i've plus than 1000 events/second wich...
03 March 2015 6,499 3 View
When you do soft agar colony formation assay, do you treat (with drug/ modulator) cells growing on agar or you pre-treat the cells for a particular incubation period and then seed the treated...
03 March 2015 4,777 5 View
The Gibson Assembly Mix is assembled on ice for obvious reasons, including not chewing back those ends too far. However, is there a suggestion to go into a machine pre-warmed to 50 degrees C or...
03 March 2015 8,248 1 View
I want to perform Co-IP from the bovine kidney cell line transfected with the Myc-tagged bait and HA-tagged prey proteins. I am using Pierce RIPA lysis buffer for the cell lysis but the lysate I'm...
03 March 2015 2,791 7 View
I am treating C2C1 myotubes with free fatty acid and would like to measure mitochondrial specific super-oxide production. I will be using MitoTEMPO for specificity. I have couple of questions...
03 March 2015 9,006 17 View
I am interested to find out the composition of ILCs 3 in colon of GvHD patients. Since I only have paraffin embeded tissues, all I can do is staining cells and taking to microscope. It would be...
03 March 2015 273 6 View
We tried several times to isolate RNA from HK-2 cells with the qiagen RNeasy kit (with RLT buffer). We can do the isolation of RNA (with Qiagen kit) from other lines of human immortalized proximal...
03 March 2015 7,247 2 View
I need to check for the MLR (Mixed lymphocyte reaction) for the particular cells using MTT assay. I have to co-culture the responder cells (CD8 T cells) from one mouse strain with the stimulator...
03 March 2015 6,964 15 View
Hi I am trying to do double labeling of endothelial cells in FFPE tissues by using a Biotin conjugated reagent against a surface marker, and an indirect fluorescence approach (primary Ab and...
03 March 2015 8,585 15 View
Dear colleagues, I am trying to determine mitochondrial membrane potential with the help of TMRE but it's not working. I am following the protocol from abcam website which is incubating the cells...
03 March 2015 6,038 5 View
Hello I’m working on cell adhesion assay on fibronectin coating. I have prepared aliquote of fibronectin (50µg/ml of steril water) freezed at -20°C. However, when I thawed my aliquot, there was...
03 March 2015 6,379 5 View
I was using 10th day embryo, DMEM 450 ml, 10% FBS 50 ml, streptomycin 5 ml, tripsin for CEM culture. Three times I failed due to contamination. Finally, I got good cell growth in all the six well...
03 March 2015 2,187 4 View
Which viability assays can I do to confirm chondrogenic differentiation from MSC pellets? Cells were pelleted in micro centrifuge tubes and differentiated.
03 March 2015 6,303 7 View