What is the transfer condition for handmade 1.5mm 15% gel to transfer histone H4?

I AM USING 20% METHANOL IN TRIS GLYCINE , 0.2 MICRON NITROCELLULOSE MEMBRANE FOR 100 V HR . the ponceau staining looks weaker compared to precast gel 10-12% with the transfer using sds and...

02 March 2018 327 3 View

Does molten magensium react with nickel curcible?

08 September 2014 7,456 4 View

Can we store Annexin V FITC stained cells and use after a week?

Can we stain the cells with annexin v fitc and fix with pfa and then store? If yes, can anyone suggest the optimized protocol for flow cytometry?

03 April 2014 8,449 6 View

Does the primary Ab and secondary Ab from different companies matter?

I am using cell signalling primary antibody and secondary antibody from sigma and substrate is NBT/BCIP. primary Ab dilution 1:1000 (3hrs rocking) sec Ab 1:10000 (45min rocking) primary ab...

02 March 2014 1,594 6 View

How do you add output styles in Endnote web?

I want to add the XXX journal output style in endnote web. In my endnote web only 21 output style are available, e.g. numbered, Ris, science, etc.

02 March 2014 6,054 12 View

What might be the reason for ghost bands in western blot?

What might be the reason for ghost bands in western blot? 1. I have used 0.5% skim milk overnight blocking 2. Primary antibody dilution 1:1000 3hrs rocking (3xwash PBST/ PBS) RT 3.Secondary...

01 February 2014 6,593 6 View

Why are bands are not visible in PVDF membrane after being stained with ponceau?

I have transferred gel in PVDF membrane and stained with ponceau but I couldn't see any bands on the membrane, instead, the whole PVDF membrane is turned into pink color and the color is not going...

11 December 2013 2,417 3 View

What is the difference between scrapping and trypsinising cells for extracting protein?

Is there any difference in protein content or alteration in phosphorylation of protein by using scrapping method and trypsin method for doing western blot?

10 November 2013 354 9 View

Why protein from SDS PAGE is not transferred onto nitrocellulose membrane?

While I'm transferring the protein from SDS gel to Nitrocellulose membrane only marker is getting transferred but not protein. I used 100v for 1hr to transfer. Can we reuse a transfer buffer? Is...

08 September 2013 8,118 21 View

Can we reuse a primary antibody which is diluted skimmed milk powder? If yes, can anyone suggest storage conditions?

-

01 February 2013 6,862 9 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

Can two EC50 values with different hill slopes be compared?

Hello, I fit a classical dose response curve Y=Bottom + (Top-Bottom)/(1+10^((LogEC-X)*HillSlope)) to my data (not related to inhibitors etc). The model I use is purely qualitative but it fits...

23 February 2021 8,155 5 View

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...

17 February 2021 4,844 4 View

What is the best temperature to coat an ELISA plate?

Hello, I want to control the temperature of my ELISA plate during incubations to get consistent results. In general, what is the best temperature to coat the plate with antibody? I have been...

16 February 2021 2,377 7 View

Is there a way to tell based on appearance which protein crystal should diffract the best?

I have had unheard of success with protein crystallography lately from a super successful protein expression and purification batch. I have attained a lately reproducible vast amount of crystals...

09 February 2021 5,155 5 View

How to measure the concentration of a labeled protein with no tryptophan or tyrosines?

Hey I'm trying to measure the concentration of a protein I've purified and then labeled with Atto 647 flourescent dye. The protein has no tryptophans or tyrosines (although does have...

04 February 2021 4,482 4 View

How to solve error "Cannot find position restraint file restraint.gro (option -r)." on GROMACS?

Hi, I am about to do energy minimization but the above error occurred, I checked my .mdp file and .top file but could not find the problem. TOP file ; Include forcefield parameters #include...

01 February 2021 780 3 View

Why is Fermented Plant Juice (FPJ) always diluted with water? Could it be diluted with other solvents such as rice water?

I am currently making a research paper with my classmates in Philippine Science High School and we are trying to find references that mixed Fermented Plant Juice with other solvents instead of...

17 January 2021 5,099 4 View

Any stable dye to color acrylamide hydrogel beads?

Hi! We are generating hydrogel beads(50-60um) using microfluidics as single-cell beads. To simply all the experiments (especially the centrifuge), I am looking for a dye. The dye should be stable...

14 January 2021 4,383 8 View

How to phosphorylate specific residues on pymol?

Hi everyone, I am using the pyTMs plug in on Pymol to phosphorylate a particular threonine in my protein. I am having difficulty in selecting only one or two residues - it seems I can only...

05 January 2021 230 3 View