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Questions related from Duy Le Pham
Hello all, I accidentally found that adding bovine serum albumin (BSA) in serum-free RPMI 1640 medium (1% w/v) caused A549 cell detachment. Have you known about that effect of BSA? I tried to find...
06 June 2016 9,725 2 View
Hello all, Could you please let me know whether FBS could inhibit the degranulation of eosinophils, mast cells and neutrophils? As I saw in some reports, FBS could inhibit eosinophil...
12 December 2015 9,649 3 View
Hello all, I am using MTT assay to test the toxicity effect of a substance on A549 cells (adherent cell). I seed 5x10^4 cells/well of a 96 well-plate, incubate overnight for cell attachment. Then...
06 June 2015 7,041 10 View
Hello all, I am trying to induce NET induction by IL-8 (100 ng/ml) using healthy neutrophils and patients' neutrophils. I quantify the NET induction by Picogreen assay. In case of healthy...
05 May 2015 8,448 4 View
Hello all, I stained the cells (LAD2) for immunofluorescent microscopy and mounted the slides by mounting medium with DAPI (Vectorshield). When mounting, I pressed the cover slips against the...
09 September 2014 9,208 2 View
I have a MISSION shRNA plasmid DNA provided by Sigma (please see the information in the attched file). I transformed the plasmid into TOP10 Ecoli strain by heat-shock method. After growing on agar...
09 September 2014 3,866 11 View
Hello all, I would like to knockdown some gene in LAD2 cells to produce a stable expressing cell line. Could you suggest me an appropriate method to do? Because LAD2 cells are stem cells and they...
06 June 2014 1,039 0 View
I am using 80V/90min to transferring LC3 from 12% SDS-PAGE to PVDF membrane. But recently, I have not gotten any band of LC3. As I've read somewhere that LC3 could swim through the membrane if we...
05 May 2014 275 13 View
When I performed WB with cell lysate from human neutrophils, I got 2 bands of beta-actin. For other cell lines (HMC-1, LAD2 cells) I just only got 1 band for b-actin. I used the same antibody and...
04 April 2014 3,087 0 View
I used to performed Western Blot to detect a protein with #14-16 kDa molecular weight (LC3) a few months ago. It worked very well. I have stopped doing that experiment for one month. I am now...
03 March 2014 6,789 10 View
Hi all, Can Skimmilk be used as blocking buffer for ELISA tests with human serum samples? Can anti-milk-IgG or IgE antibody in serum samples interacts with skimmilk that blocks the plate? Thanks...
02 February 2014 9,484 18 View
I am working with LAD-2 cells (a human mast cell line). I usually perform hemidepletion every 7 days. Within this period, I check the cells every 2-3 days and pipette up and down (with slow...
02 February 2014 8,585 14 View
I would like to treat A549 cells with IL-1beta and then collect the supernatant for measurement of IL-18 level using ELISA. I had some confusion as below: 1. Is A549 a source of IL-18...
12 December 2013 6,626 0 View
I am trying to clone 2 fragments of 1.6 and 1.1kbp into pGL3 basic vector (4.8kbp) using In-fusion HD cloning kit (Clontech). I added totally 90ng of 1.6kbp fragment and 70ng of 1.1kbp fragment...
08 August 2013 3,687 12 View
I am going to culture the airway epithelial cell line A549. After taking the vial out from liquid nitrogen, we have to incubate the vial in water bath at 37oC. Why do we set up the temperature at...
04 April 2013 4,689 11 View
I did ligation steps to get pGL3-basic vector with the insert, then transformed into competent cells and cultured on an agar plate with ampicillin. After incubating, I had to choose colonies for...
03 March 2013 2,053 16 View
