Can I reuse western blot transfer buffer?

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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

Dear all, I was wondering if anyone knows whether MGLtools and Autodock can run on Mac OS?

I have tried to download the 64-bit version of MGLtools however, as many times as I have downloaded and uninstall the programs, an error arises that the app needs to be updated. Thank you.

03 March 2021 5,997 2 View

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

I got 02 moderator values defining Johnson-Neyman significance regions. Is it an error?

Hi everyone, I am conducting research for my Master's thesis. I am using PROCESS by Johnson-Neyman to analyze my Moderator model. I test the relationship between Public Service Motivation and...

03 March 2021 2,350 2 View

Do I need to do linear regression before running mediation analysis using PROCESS MACRO by Hayes ?

I just wanted to check if I need to run a linear regression separately if I am using PROCESS MACRO to run mediation analysis. Thank you.

02 March 2021 4,359 3 View

Do I need to strip the PVDF membrane before staining it with colloidal gold?

Dear Colleagues, After running Western blot on PVDF membrane and detection using ECL, I would like to stain my PVDF with colloidal gold to be able to allign the ECL image with total proteins on...

02 March 2021 7,829 3 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 View

What SPSS analysis should I run with one IV (with two levels) and two DVs?

Hi, I am trying to figure out which test would be most appropriate to run for the following question: Is there a difference in task blood pressure between smokers and non-smokers? There are two...

01 March 2021 8,235 2 View

OpenBCI eeg--has anyone used them?

I'm looking to try out an off-the-shelf EEG set for some exploratory work. Curious if anyone has used the OpenBCI, and how much set-up work is required. I've heard they are more difficult to get...

01 March 2021 4,128 3 View