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

What is the function of XBP buffer in exosome isolation?

04 May 2017 1,036 2 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

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

What is the density for pva electrospining?

I'm Juan Carlos, a high school student who is working with electropinning, I made an electropinning machine with a 21g needle and a 30kv high voltage source and the distance between my needle is...

28 February 2021 298 3 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

Is that possible to do an impedance measurement without FRA module?

One of my colleagues said, "the impedance you can measure with the three electrodes and with the specific sequence varying the frequency" Is it possible to configure the potenciostat without the...

21 February 2021 418 2 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

Choice of Ab for Co-IP: Does an anti-(tag) antibody (such as anti-flag/ anti-myc) a must if I am using co-transfected cells for Co-IP?

Hi guys, I am trying to detect the binding of 2 proteins by Co-immunoprecipitation. However, the choice of antibody is confusing me. I tried probing the endogenous protein right away but the...

16 February 2021 9,050 5 View

How to show in a scientific paper format the post-hoc test of 4 variables involved, when 3 by 3 variables are significant?

In a study involving 4 variables (species(2x), temperature(2x), nutrients(2x) and light(3x)), 3 by 3 variables, for example, "species", "temperature" and "nutrients" are significant. I end up with...

15 February 2021 8,018 2 View

Kinetics of Enzyme production in Recombinant E Coli?

Hello, I am trying to find kinetic information (such as rate of enzyme production etc.) on the expression of taq polymerase in E-Coli after inserting a recombinant plasmid. Any help would be...

12 February 2021 1,386 3 View

Is it possible to quantify recombinant protein in the supernatant using SDS PAGE ?

I would like to quantify a recombinant protein (with native signal sequence) secreted by Pichia pastoris using the SDS PAGE. Since my standard protein (plant origin) is not pure, I would like to...

07 February 2021 3,880 9 View