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...

03 June 2019 9,926 10 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 &...

15 January 2019 3,876 4 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...

13 November 2018 5,368 7 View

QPCR spike in normalization?

Can someone give me an easy example of how to normalize qPCR data against spike-in control? Thanks

05 March 2018 9,534 1 View

Hemolymph extraction pupa stage?

18 October 2016 4,507 2 View

Is there an efficient way to calculate rate constants for vibrational energy transfer in CO2-CO2 collisions, especially near the dissociation limit?

Dear all, I am currently modeling the plasma kinetics in pure CO2 plasmas with a large focus on vibrational kinetics. The plan is to optimize the vibrational kinetics (VV and VT transfer), if...

24 April 2016 7,217 4 View

Does anyone have a good protocol for colony PCR on Stellar competent cells?

I am screening for correct clones after an In-Fusion cloning and transfection in Stellar competent cells. I usually pick up a colony and add it to the PCR-mix (I use HotStart polymerase with an...

01 December 2013 8,550 7 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