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Questions related to Cellular Biology Techniques
I know that in human cell-lines, some of the N-linked sugars that are added to proteins during maturation in the secretory pathway contain sialic-acid residues. This gives the N-linked sugars a...
07 July 2015 692 4 View
I am treating AR42J cells with 10-8 M cholecystokinin (CCK) for different time intervals up to 8 hours. Following this, I need to check for markers of cell death and for activation of the Unfolded...
07 July 2015 7,469 4 View
Hello, I try to establish a 107 a assay unfortunately without success. I used human PBMCs and cocultured them with several cancer cells (with and without developed drugs). The flow results looked...
07 July 2015 1,277 1 View
I'm curious as to how to terminate protein expression in T7 Express Competent E. coli (high efficiency) after gene induction with 0.2% arabinose? Is there any way to halt this besides waiting for...
07 July 2015 4,450 3 View
The resistance reading from WPI EVOM2 is ohm x cm2 or just ohm? I read the manual very carefully and thought it's ohm x cm2 and we don't need to convert it. But the TER value I got for my RPE...
12 June 2015 5,327 2 View
How Type I IFN involvement of oxidative stress induce the DNA damage response of aging process?
06 June 2015 4,112 2 View
I am trying to detect stimulated Tregs and their IL-10 expression in cultured PBMCs (not isolated from the cells mixture) by flow cytometry. For other T cells phenotyping and evaluating of...
06 June 2015 9,992 10 View
I am trying to do clonogenic assay with HCt116, Ht29 and Panc1 cell lines. I did plating with different cell concentrations for HCT116 (200, 300, 500 and 1000/well in 6-well plates). I had good...
06 June 2015 3,824 13 View
I am studying the secretion of the gut hormone (GLP-1) using the NCI-H716 cell line. Since it is my first time to use this cell line in particuljsr and suspension cell line generally, I am...
06 June 2015 6,182 3 View
I want to use, 1:10 MOI, (cell: bacteria). Assuming that one well of cell is 1 x 10^5, thus, the no of bacteria per well should be 10 x 10^5. The bacteria is V. cholerae, and from my reading, at...
06 June 2015 839 1 View
Does anyone have experience with the CellROX Green reagent for oxidative stress detection? We are trying to detect oxidative stress in cells(live or fixed) in 35mm plastic dish. Unexpectedly, the...
06 June 2015 8,785 13 View
Hi! I am trying to reversibly inhibit DNA replication of my mouse cells. I read that a lot of inhibitors actually cause DNA crosslinking and eventually lead to apoptosis, which I don't want. I am...
06 June 2015 5,154 9 View
I use regular KGluc solution, but with ajusted to 450 mOsm osmolarity. Bath solution is regular Tyrode-based, with normal osmolarity. Under these conditions gigaseal forms very quickly, but when I...
06 June 2015 6,858 4 View
I found a paper (ref below) that describes uptake of ADP-ribose (3H-labelled) in erythrocytes. I was wondering if anyone has any experience with this protocol or has looked at ADP-ribose uptake in...
06 June 2015 8,144 1 View
06 June 2015 5,245 2 View
I am a student researcher, investigating whether glibenclamide can restore insulin secretion in cells with Wolfram syndrome (WFS1 knockdown). The team I am working with have been gifted some MIN6...
06 June 2015 2,041 3 View
I've been using the silicon oil filtration method from the Heldt papers, which uses tritiated water and tritiated sorbitol to measure intracellular volume. The scintillation counter reads...
06 June 2015 6,073 0 View
To calculate growth index, I am using the below formula for Dry weight of the cells. W (final)- W(initial)/ W(initial) However, the resulting number is high almost higher than five. I was...
06 June 2015 4,034 1 View
Hi, I'm currently trying to detect TNF via IF in mouse leukocytes (specifically neutrophils but will take what I can get at the moment). I have tried a range of positive controls (splenocytes...
06 June 2015 4,642 3 View
it is well known that mutations in some genes can leads to cancer condition. if in that case how those mutated gene products can able to participate in different pathways? especially in the case...
06 June 2015 4,641 4 View
Searching the web i found that Zymo Research makes a kit for making competent cells and transforming them in aprox 20 seconds without incubating the mix on ice for 30 minutes, without heat-shock...
05 May 2015 3,644 4 View
Currently, I am trying to stain muscle frozen section with three antibodies CD31, PAX7 and laminin. I have tried fixing them using ice-cold 100% methanol and 4% formaldehyde. block for 45 min and...
05 May 2015 5,941 14 View
I'm interested in the functional impacts of Golgi fragmentation - what are good recent articles to read that discuss Golgi structure/function?
05 May 2015 4,735 3 View
Hey everyone! i am trying to analyze the effect of a drug on mammalian cells. I know that I have to starve the cells before treatment but someone told me that I have to starve the cells again...
05 May 2015 7,907 5 View