Immunoprecipitation: beads without antibody binding still give a signal?

Hi all, As a negative control, I've used a set-up in which: 1) beads without antibody binding were incubated with the sample and then put on WB. I don't see any heavy chain (which is logic),...

01 February 2020 7,255 2 View

Slower migration on SDS-page in complex sample?

Hi all, Is it possible that the migration of my protein is delayed due to the complexity of the sample? When I run a western blot with 1) a protein extract from locust tissue and 2) hemolymph...

05 June 2019 2,727 11 View

Secondary antibody gives same signal as primary?

Hi, After immunoprecipitation and elution, I check the eluate with western blot. If I incubate the blot ONLY with secondary antibody, it gives the same signal as incubatin with primary &...

31 December 2018 1,079 5 View

Western blot on immunoprecipitation eluate detects related protein?

Hello, So, first I perform immunoprecipitation. The eluate is then subjected to immunoblot analysis using an antibody against a related protein (equal size) and gives a band. How sure am I the...

10 November 2018 4,435 8 View

Allways same (wrong) sequence after sanger sequencing?

Hi, I've cloned and sequenced my PCR product but, besides a good run, I allways find the same (wrong) sequence in other runs. Running a BLAST gave no result. What could this be?? Thanks!

01 February 2018 3,433 0 View

Agilent Bioanalyzer miRNAs of 35nt?

Hi, My sample contains miRNAs, but when I use the small RNA chip (Bioanalyzer Agilent), I get a peak at around 35 nt instead of 22 nt? I'm pretty sure that there are miRNAs in there; any...

10 November 2017 7,578 4 View

Only cds on NCBI?

Hi, When you look up an mRNA seq of a certain gene, it says "complete cds" on NCBI. This means that only the coding sequence is given, right? When you compare this mRNA seq to a NCBI TSA...

08 September 2017 7,470 2 View

How to separate a complex fluid in a vesicular and non vesicular part

Hello, Is it possible to separate a complex fluid in a vesicular part (containing vesicles / exosomes) and a non - vesicular part, containing other proteins? Many thanks!

05 June 2017 2,792 4 View

Synthesis of RNA mimics/antagomirs?

Hello, I was wondering how RNA mimics and antagomirs are being synthesized. I don't need the details (volumes etc), but just a general overview of enzymes being used etc? Thanks! Cheers

09 October 2016 2,398 1 View

Different elution methods for immunoprecipitation result in different bands on western blot?

01 January 1970 5,929 6 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

What is the minimum number of exosomes/ml to have a good yield of total RNA for Illumina sequencing?

I am using IZON qEV2 columns to isolate exosomes from 100 ml of cancer cell conditioned medium. in NTA analysis I usually see concentration of 2x10^9 particles/ml. But when I proceed with this...

20 February 2021 6,879 3 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