How many RNAse H I need to add to the hibrid DNA (80 pb)- RNA (21 pb) 20 mM in 200 uL of Tris 100 mM pH8 and Mg2+ 10 mM T=35 C?

The idea of my project is: I ve made a sytem of one DNA which hybirizes with tho DNA with Cy3 and Cy5 each one (initial complex), when I add a fourth sequence of DNA at pH6 a triplex structure...

04 May 2016 2,504 0 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

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23 February 2021 4,014 3 View

How to reduce static electricity of powder after lyophilization?

I am working on synthesizing and then purifying peptides using the SPPS system. After purification of the peptide, the peptide powder is obtained using a freeze dryer. When measuring the powder,...

22 February 2021 10,099 5 View

Can we remove the fmoc group after peptide cleavage?

I have synthesized the peptide using Fmoc based solid phase peptide synthesis. Also I have cleaved the peptide from the resin and lyophilized it, but later realized that, I forgot to deprotect the...

21 February 2021 4,537 3 View

How T cells recognise non-peptide antigens?

This might sound very basic question, but please share with me an explanation of how the T cell is activated when the antigen is not a peptide? What happens? And what are the molecules involved in...

21 February 2021 564 3 View

Help with Amyloid beta detection protocol (western-blotting)?

Hi everyone, I am a first year PhD student working on amyloidosis. I would like some tips and tricks to crack efficiently my current nightmare, i.e. the western blotting of the Amyloid beta and...

11 February 2021 2,513 3 View

How to assing secondary structure to calculated peptide/protein having xyz file?

Hi, I have an optimized structure of some peptides and I would like to represent it as a secondary structure - cartoon. Ok, so server apps ask for pdb file (like Stride). I tried in different...

04 February 2021 517 3 View

Identical spectra for L- and D-peptides after they self-assemble?

Hi dear all, I need advice. I have two peptides with the same sequence but one consists of L-amino acids only and the other consists of D-amino acids only. when they self-assemble in aqueous...

03 February 2021 2,032 3 View

FLAG purification, elution doesn't have protein?

I am purifying a heterotetramer, with one subunit having a 1D4 tag and the other having a FLAG tag. I purified from Sf9 with HBS pH7.3 and 0.002%LMNG first with Anti-1D4 resin and got an adequate...

02 February 2021 3,860 1 View

How long can synthesized peptides using SPPS be stored? Storage Conditions?

I have been synthesizing peptides on resin beads. I am wondering what best storage method is there to store the beads after synthesis before cleaving. And how long can they be stored in that...

02 February 2021 5,636 5 View