May antibodies used for immunofluorescence be reused?

Hello, I am wondering if it is alright to reuse my antibodies for immunofluorescence experiments. I have reused primary antibodies with success for the first two experiments, but then I saw a...

01 February 2020 6,113 3 View

Microwaving food: good enough for humans but not cells in culture?

Hello, Recently I asked my colleague whether we may simply microwave culture media for mammalian cell culture in order to warm it prior to use. He told me that we may not microwave cell...

11 December 2019 7,405 2 View

In immunofluorescence staining, will the presence of phosphate in buffers affect the phosphorylation status of proteins of interest?

Hello, I am attempting immunofluorescence of a membrane-bound, phosphorylated protein. I typically use PBS in my blocking buffer. I also use PBS and PBST to wash. I have been largely...

10 November 2019 1,063 1 View

Is it possible to over-stain with DAPI?

Hello, I am performing immunofluorescence of HepG2 cells. I believe that some of my stocks may be contaminated with Mycoplasma because when I stain them, their membranes and portions of their...

10 November 2019 515 3 View

Should I use a protease/phosphatase inhibitor when conducting immunofluorescence?

Hello, I am attempting to detect a phosphorylated protein using immunofluorescence. Should I include a phosphatase inhibitor in my blocking buffer? Will a protein's phosphorylation status...

09 October 2019 7,749 1 View

Mysterious dancing dots: bacteria, viruses, or Brownian motion?

Hello, Recently I performed lentiviral transduction on several cell lines. On day one with the lentivirus, I observed black aggregates moving (dancing, vibrating) around my cells. These are...

01 February 2019 3,417 4 View

What is the impact of pouring bleach down the drain?

Hello, When utilizing bleach for biological applications, is it advisable to pour excess down the drain? What is the ecological impact of bleach entering the septic system? Is bleach...

01 February 2019 213 4 View

Can a human survive on DMEM?

Hypothetically, could someone survive by consuming DMEM with L-glutamine? I understand this is not recommended. I just wonder, because DMEM is formulated for human cells. Thank you.

01 February 2019 8,839 4 View

Can one see Brownian motion under a light microscope?

Hello, Is it possible to observe Brownian motion of particles with a light microscope at 40X? I am asking for reasons relating to mammalian cell culture. Thank you.

01 February 2019 1,155 3 View

Can methanol be used in place of ethanol for microbiology?

In a pinch, can 70% methanol replace 70% ethanol for sanitizing surfaces prior to cell culture?

31 December 2018 3,070 13 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

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02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View