18 Questions 18 Answers 0 Followers
Questions related from Raheleh Tooyserkani
Hello; I perform antibody purification using protein G column, eluting the antibody in PB. Then I prepare several antibody aliquots and store them in -20 C. I use this polyclonal antibody as...
13 September 2021 8,034 3 View
Hello; I've done MTT assays to investigate whether my peptide drug could inhibit tumor cell proliferation. I used MDA-MB-231 as positive cells and MCF-7 as negative ones which do not express...
09 May 2020 753 4 View
Hello everybody! I'm trying to label some M13 phage particles with FITC. I checked some papers; I use 0.3- 1 M bicarbonate buffer (pH 9) as conjugation buffer. I prepare 1mg/ml stock solution of...
28 December 2019 4,117 2 View
Hello; I've dissolved my synthetic peptide in pure acetic acid and tried to purify it with reverse phase HPLC. I got weak peaks during analysis, however I injected at least 5 mg of my peptide!...
25 March 2019 3,696 4 View
Hello; I've treated my cells with IFN-gamma to induce PDL-1 expression. Then, I investigate whether a synthetic peptide specific to PDL-1 could bind PDL-1, through flow cytometry. To show peptide...
28 January 2019 9,979 6 View
Hello; I want to calculate Kd value for a FITC-conjugated peptide which specifically binds to a cell receptor. I've studied some protocols, but still confused! Does anyone have any experience or...
03 December 2018 3,711 2 View
Hello everyone; I've isolated a 12-mer peptide against PDL-1 from a phage library. During in silico analysis I realized that my peptide tends to bind other sites rather than desired binding site!...
28 April 2018 6,398 4 View
Hello; I use a random peptide phage library to isolate specific peptides to PDL-1. I use the commercial host strain ER2738 for phage amplification and titration and the selection system is based...
17 October 2017 6,633 0 View
Hello; I've panned a 12-mer M13 phage library against target in solution using protein A sepharose beads. following the first round, I got no blue plaque after titration and amplification even in...
17 July 2017 3,690 4 View
Hello; I need a practical protocol to conjugate protein G to magnetic beads. Does anyone have any idea about that? Thanks
18 June 2017 4,922 4 View
Hi there; I'm going to do phage panning against the protein, PDL-1. I use a commercialized peptide library displayed on M13 phages. I want to select PDL-1 binding peptides which are able to...
31 May 2017 1,982 3 View
Hello, I need to determine the appropriate dose for a peptide which aims to inhibit a protein on tumor cells. But I have no idea about how to justify the animal numbers and how to that! Does...
01 February 2017 5,325 3 View
I've transfected HEK293-T cells with pCI as a mammalian expression vector containing neomycin gene as a selectable marker. I need to know the most efficient concentration of G418 for cell...
26 April 2015 1,379 4 View
I have cloned a 25KD fragment in pet32a and I got recombinant expression in BL21. The yield of expression was very good, but suddenly it stopped expressing!I tried transforming new BL21 bacteria...
09 February 2015 4,116 1 View
I have immunized rabbits and mice with DNA and I got no antibody answer in ELISA test! I need to know is there any other method to consider the presence of immune response?! I mean I want to know...
08 February 2015 2,891 5 View
I'm working on polyclonal antibody production through genetic immunization in rabbits and after 5 month of immunization I got nothing! I need to know is there a problem with Tolerance induction...
02 January 2015 2,040 4 View
I need to study experimental methods for epitope mapping for my proposal but I can't find any comprehensive article.
17 September 2013 6,209 2 View
Articles have mentioned they used antibodies to specifically detect the dimeric form but there's no sequence difference between dimer/tetramer as far as I know!
14 August 2013 2,126 1 View
