How long can I store a loading sample after mixing with loading dye and at what temperature 4°C or -20°C for using in gel electrophoresis?

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Whether L version and D version peptidesshould have identical NMR, especially in the state of self-assembly?

Whether L version and D version peptides should have identical NMR, especially in the state of self-assembly have to have identical NMR? How to use HRMS to confirm L version or D version...

01 February 2021 6,756 4 View

Protocol for in vitro cytokine stimulation of immunized CD8+ T cells?

Hi, everyone: I want to determine whether IL-2 and IL-15 have different effects on immunized CD8+ T cell effector function. In this study, I immunize WT mice with peptide on day 0 and 7, and...

02 December 2020 9,413 3 View

Do I need to run a factor analysis on all the items used in the SEM to solve the multicollinearity issues?

Respected researchers, I conducted a SEM including 5 variables (two independent variables , two mediators, and a dependent variables), the correlations between these variables range vary from...

01 December 2020 1,338 5 View

How to extract RNA from small number (100-150 at most) of sorted cells?

I'm trying to test expression of a gene in sorted cells. I plan to extract RNA from sorted cells and to do RT-PCR (reverse transcription PCR). The problem is that from sorting, I can only collect...

21 November 2020 5,467 2 View

After place the crudes on an column spectroscopy (sorry, column chromotograph), is it approprate to touch or move the crude sample anymore?

As the title has asked. For instence, using metal stick or long glass tube to get rid of the bubbles in columns.

05 November 2020 5,946 4 View

B cell isolation from mice spleen using Meltinyi negative kit?

Hi, guys, Do you have problem using B cell negative selection kit form Meltinyi? Sometime the B cell is pure, CD19+>95%, but sometimes it lower than 90%.

19 October 2020 5,160 3 View

How to separate exosome and subcellular organelle (eg. ER) of mouse liver?

I want to do tissue subcellular fractionation of mouse liver. According to the method, the speed to separate ER is over 200,000g for 120 min (Song et al., 2006). However, liver exosome isolation...

12 October 2020 4,929 3 View

How to set up rheological model (CA and CAM) ?

how to set up asphalt rheological model (CA and CAM) ? How to build EXCEL?

23 September 2020 7,692 2 View

Should 1:1 binding model be used for antibody binding in biacore(SPR)?

Hi anyone experienced in studying antibody-antigen binding with biacore (SPR) know should 1:1 binding model be used in kinetics and affinity analysis? Or should we use 1:2(antibody:antigen)? Thanks!

26 August 2020 4,130 3 View

How can we deal with the impacts of water management (e.g. drainage) when doing long-term hydrology and water quality modelling?

The anthropogenic pratices are getting common these days and many catchments experience water managements e.g., drainage, which has huge impacts on both discharge and water quality dynamics. So...

13 August 2020 1,801 11 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

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

Why are my bands weird in this Agarose gel?

Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.

01 March 2021 9,952 3 View

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 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

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 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

Nageswar Reddy Manchala

I suugest you to sotre the samples at 4degree.

Alejandro Martin

It depends on the type of sample, the type of electrophoresis, etc. I would never store RNA samples to be used for Northern blotting, but would not hesitate to do it for protein samples in denaturing & reducing SDS-PAGE sample buffer.

Bassem Refaat

I suggest -80 for RNA and -20 for both DNA and proteins.