What is the best western blot apparatus for electrophoresis of proteins?

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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

Protein-aptamer gel electrophoresis help, please??

Hi, I have problems with running gel electrophoresis. I have tried agarose gel electrophoresis and native PAGE. I have two proteins, which have molecular weights of ~30kDa and ~180kDa and two...

03 March 2021 4,275 4 View

How to analyze a particular region of my simulation box in GROMACS?

Hi, I am using GROMACS 5.1.4 to simulate water in contact with a surface. I am only interested to calculate some properties of water near the surface. Let's say at a distance of x nm from the...

03 March 2021 9,989 2 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

How to regulate SSCP temperature in simple PAGE?

I'd like to perform single-strand conformation polymorphism (SSCP) in my thesis, however I cannot control the temperature of the vertical PAGE since we are using the conventional tanks. Is there a...

02 March 2021 9,157 1 View

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

Brian Bower

I've been very happy with Biorad's product lines for both electrophoresis and transfer. They often offer a combo set that contains everything needed to begin running either Criterion or mini-gels.

Khalid Dar

well biorad is most trusted. can anybody intimate which product of biorad's wet blot is best suited and contains all requirements from electrophoresis to transfer