12 December 2019 3 5K Report

Hi. I'm analyzing the kinetics of antibody antigen interaction. I have some that show very low Koff. The problem is in the dissociation phase the curve is not only stationary but increase....the reviewer need explanation. Is a question of normalization with the base-line? I mean there are not many things I can have done a wrong

in the experiment. Please I need a acceptable justification.

More Alexander Bontempo's questions See All
Can I complement data I used in a previous publication?

I compared 4 protein trough a methodology. In a previous publication I just showed the data for 3 of them. Now I want to publish the complete set as my interest is to present the 4th protein that...

31 December 2019 7,246 2 View

Why when i do a BN-PAGE of a single SEC peak fraction i get many bands?

hi, i'm doing a protein purification and i'm wondering what are the factors make a single SEC peak give many bands in a BN-PAGE.

08 September 2019 7,503 8 View

Is a expression plasmid for HIV env protein a BSL2 agent?

Hi guys. I'm facing a institutional divergence about the BSL of a expression plasmid carrying the HIV env gp160 protein. They are make trouble believing it is a bsl 2 agent. Is that true? If is...

06 July 2019 8,134 4 View

How can i estimate different peaks in a overlaping muly-peak graph?

Hi, i made a sec an the chromatogram is composed of overlaped distributions(peaks). How can i estimate the apportation of each singular poblation? Wich is the best software that also permit a...

10 November 2018 8,915 0 View

Why my mini prep is working but no my maxi considering same bacterial culture?

Hi i have a quiz for you guys. i don't know how to solve this trick. attached you ll find flow chart with my issue in dettails. Briefly, i tried to purify a plasmid trough maxiprep (two different...

10 November 2018 6,866 5 View

Nobody has a detailed protocol for CHO transfection with PEI?

Hi, this is my first time i try protein expression so i need a really complete protocol to perform CHO-K1 transfection using PEI scalable up to one liter culture. I need a protein expression for...

04 May 2018 2,314 3 View

Why hiv gp140 trimers in chromatography shown a peak between ferritin and thyroglobulin references?

I saw some gromatography graphs and i can't understand why gp140 trimers who should weight 412kDa shows the peak between ferritin (440kDa) and thyroglobulin (669kDa) references. Thank you.

31 December 2017 1,091 2 View

Similar questions and discussions
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

Have someone successfully used the ER alpha (Santa Cruz sc-8002) antibody in immunostaining ?

Santa Cruz now recommends this new antibody (F-10) but I can't find a publication using this reference. I'm working on human et non human primate hypothalamus and so far, it doesn't work. Is...

02 March 2021 10,003 2 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Does anyone know an alternative product for cardiac Troponin T antibody for Immunofluorescence (Thermo # MA512960)?

We have been using Thermo Fisher anti-cardiac Troponin T antibody for immunofluorescence staining in FFPE mouse heart sections but it was recently discontinued. The literature shows it to be the...

01 March 2021 4,937 1 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

How does an antibody have full intact antibody + half antibody?

Hello, I was just wondering how does a antibody have full intact antibody + half antibody? Is it because of the drug conjugation?

28 February 2021 1,577 1 View

Does anyone use antinuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples?

I am looking for anti-nuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples. Any suggestions highly appreciated. Thank you.

28 February 2021 6,512 3 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

Anyone know of a good progesterone antibody?

I am looking for a good progesterone antibody to quantify levels of progesterone in zebrafish. Applications would be ELISA, and immunohistochemistry.

28 February 2021 5,929 2 View

How to calculate Ligand-active site distance in a ligand-protein complex from MD simulation?

I have run a MD simulation of a ligand-protein complex in GROMACS using CHARMM forcefield. Reviewer has asked me to calculate Ligand-activesite distance and binding free energy of ligand with...

27 February 2021 4,703 1 View

Our partners will collect data and use cookies for ad personalization and measurement. Learn how we and our ad partner Google, collect and use data. Agree & close