Is there a limit on the protein size?
Which parameters should I have in mind to choose one or the other?
How to quantify recombinant expressed protein if some is secreted to medium and some gets attached to the membrane? (also some inside the bacteria but I'm not interested in this fraction). I...
06 July 2013 9,435 9 View
Is there any online site where you can put the sequence and search if it has sites where you can use universal primers (T7 or M13)?
06 July 2013 1,107 9 View
I did a little test to see if my substrate and conjugated Ig were ok. When I added 1ul of anti Ig-Y (secondary conjugated antibody) directly to 1ml of my TMB working solution it turned orange...
06 July 2013 3,507 5 View
Which other reagent can I use ?
05 June 2013 7,267 7 View
I´m using as secondary antibody: HRP Conjugated anti chicken IgY (http://www.genetex.com/Chicken-IgY-HL-antibody-HRP-GTX26753.html) and TMB substrate...
05 June 2013 705 10 View
I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...
07 August 2024 505 3 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,989 3 View
Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC
01 August 2024 967 3 View
Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...
31 July 2024 9,892 4 View
Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....
29 July 2024 5,007 0 View
I want to do 2,3-butanediol dehydrogenase(BDH) enzyme purification to confirm its activity for 2,3-butanediol. Before that, I need to confirm which N or C terminal tagging is better for enzyme...
28 July 2024 366 3 View
I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...
28 July 2024 3,178 3 View
I'm having problems with ion exchange chromatography. After applying my sample (a fluorescent protein), a preferential path inevitably forms. The matrix I use is DEAE-Sephacel. Could someone...
27 July 2024 6,605 0 View
It has been long known that EDC carbodiimide and NHS easter's half life agains hydrolysis are highly related to pH. However, very surprisingly, not yet found a paper giving a table/chart of their...
25 July 2024 8,738 0 View
I use 0.5% sarkosyl to solubilise recombinant proteins contained in Inclusion bodies of E. coli. sarkosyl very effeciently solubilizes proteins from Inclusion bodies and I am trying to use...
25 July 2024 5,001 2 View