Is there a "good" transDUCTION protocol?

Hello dear colleagues, I am experiencing strange problems when performing transDUCTION on my CD4/CD8 cells (separated using Miltenyi beads from total PBMCs from a buffy coat). For now on it is...

03 April 2016 6,843 1 View

Any limitations for expression in T-lymphocytes?

Hello everyone, we are trying to transduce T-lymphocytes (TLs) with vector containing IRES, and cannot see any expression using flow cytometry (our target protein is conjugated with GFP). The...

02 March 2016 6,562 4 View

Is it possible to overcome co-expression?

Hello, in my expression cassette I already have 2 proteins that are expressed using IRES. However I'd like to express another protein at the same time (a homing molecule), but, honestly, I do not...

01 February 2016 5,983 3 View

A good biotin conjugate for FACS?

01 February 2016 8,243 5 View

What's the best solution for T cells expansion?

Hi, I am trying to perform an expansion of T-cells, purified (by miltenyi columns) from PBMCs and transduced with construct of interest. I am wondering if there's a reliable and "working" protocol...

11 December 2015 386 5 View

Best lab journal software?

Hello, everyone. We all know how, sometimes, it's difficult to write down everything into a lab journal, while I believe there are other approaches apart MS Word :) Would anyone be able to share...

06 July 2015 5,357 0 View

Can anyone suggest a positive control for Raji cells?

05 June 2015 9,775 3 View

What's the "normal" percentage range for PAN-T cells out of PBMCs?

Hi, colleagues. Unfortunately I have just started to work in this new field for me, PBMC purification from blood bags, MACS separation etc. So what I do is out of a blood bag from hospital I do a...

05 June 2015 5,812 8 View

Do I need to preculture cell for siRNA experiment?

Hello, I am planning to perform a siRNA knockdown using lifetechnologies Lipofectamine RNAiMAX reagent, so taking into account they require 60-80% confluence of cells on the moment of transfection...

02 March 2015 1,424 5 View

FACS markers for Jurkat analysis?

Hello, I am about to complete my FACS training so I need to bring some cells labeled with different markers (correct me if I'm wrong in terms). What I have here in culture are Jurkat cells, so I...

01 February 2015 7,338 3 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View

Can I use a protein-devoid condition as my negative control in ELISA?

Hello, I am running a sandwich-type ELISA where I am trying to inhibit a protein-protein interaction using small molecules. For each experiment, I determine my Z' (Z-prime) to determine the...

23 February 2021 7,429 1 View

How to build fluid structure interaction simulation for elastic ring squeeze film damper in COMSOL?

How to build fluid structure interaction simulation for elastic ring squeeze film damper in COMSOL?

22 February 2021 5,408 1 View

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...

17 February 2021 4,844 4 View

Kinetics of Enzyme production in Recombinant E Coli?

Hello, I am trying to find kinetic information (such as rate of enzyme production etc.) on the expression of taq polymerase in E-Coli after inserting a recombinant plasmid. Any help would be...

12 February 2021 1,386 3 View

Is it possible to quantify recombinant protein in the supernatant using SDS PAGE ?

I would like to quantify a recombinant protein (with native signal sequence) secreted by Pichia pastoris using the SDS PAGE. Since my standard protein (plant origin) is not pure, I would like to...

07 February 2021 3,880 9 View

How to measure the concentration of a labeled protein with no tryptophan or tyrosines?

Hey I'm trying to measure the concentration of a protein I've purified and then labeled with Atto 647 flourescent dye. The protein has no tryptophans or tyrosines (although does have...

04 February 2021 4,482 4 View