What is the minimum concentration of glycoproteins (in total) required for analysis of N-Glycans using MALDI-TOF mass spectrometry?

I have expressed a human glycoprotein in plant ER through a signal sequence and it also contains ER retention signal at the C-terminus. Thus, if this protein contains an N-glycan moiety, it should...

11 December 2018 9,367 3 View

Should I change my PCR condition or design longer primer for low GC content (32% GC)?

I am trying to amplify a gene from plasmid DNA which has A-T rich sequence and low GC content, 32% GC. However, I could get the desired product which is around 900bp, but, I found lots of PCR...

09 October 2015 6,075 5 View

How can I optimize PCR conditions for a reverse primer of length 51 base pairs (Tm 82 Deg) and forward Primer of length 24 base pairs (Tm 72.5 Deg)?

Hello I am trying to amplify a plasmid DNA using a forward primer of 24 base pairs and reverse of 51 base pairs. In the reverse Primer among 51bp, 24 is hybridizing sequence and remaining 27bp is...

05 June 2015 5,310 7 View

How to overcome the elution of non-specific bands in protein purification by Ni-NTA resin under native condition (with 6XHis-tag)?

I have eluted my protein with 250mM imidazole and got some non specific bands in the elution and yield also not good. Besides, during washing step i was used 20 mM imidazole washing buffer and...

01 February 2014 9,489 12 View

what is the simplest definition of ion exchange capacity

What this unit identifies for the ion exchange capacity ca. 0.92 meq. g-1. I am a newcomer in the membrane area. I just started. I highly appreciate your help

08 May 2021 0 0 View

what is the simplest definition of ion exchange capacity

What this unit identifies for the ion exchange capacity ca. 0.92 meq. g-1. I am a newcomer in the membrane area. I just started. I highly appreciate your help

08 May 2021 0 0 View

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

POlymer+DMSO solution and moisture??

What could be the reason of moisture at the sides of Vial when I am preparing my polmyer+DMSO solution for membrane fabrication? What etiquettes I am possibly missing here?

28 February 2021 6,516 5 View

IF troubleshooting: looking at an intracellular protein that localizes to the cell's and how to reduce increase in signal due to cell ruffles?

I am currently trying to stain for PI3KBeta in MDA-MB-231 cells. My current issue is that 1) beta stains all over the cell and 2) there is an increase in signal at the edge of the cell and this...

24 February 2021 8,582 1 View

Cloudy/high background western blot - can anyone help me?

Hi everyone! I try to detect VDR protein (~48 kDa) using Western Blot. After SDS-PAGE using 10% MP TGX Stain-Free Gel from BioRad I transferred the protein onto an metOH activated PVDF membrane...

23 February 2021 6,266 3 View

Protocol for membrane extraction E. coli?

Hello! I am interested in determining whether a poly-unsaturated fatty acid I have engineered E. coli to produce is being incorporated into the membrane of the cells or if it is simply a free...

21 February 2021 7,447 2 View

What are the best cations for preparing competent cells, Ca2+ or Mg2+?

I have been reading various articles and most state that treatment with CaCl2 produces higher efficiency of transformation than MgCl2. Can anyone explain why Ca2+ is better than Mg2+ at...

16 February 2021 8,519 5 View

I have some problems with the mass transfer part of the CFD simulation of membrane distillation.Can anyone help me?

I am working on modeling of Direct Contact Membrane Distillation system with COMSOL Multiphysics software. I have some problems with the mass transfer part of the simulation because I don't know...

14 February 2021 379 2 View

Regarding immunology, transmissivity, herd immunity and vaccines in Sars CoV-2?

This is a follow-up on an earlier question regarding mechanistics of vaccines: why current vaccines don't block transmission. In summary, they don't engage mucosal immune cells to produce IgA....

08 February 2021 8,321 3 View