44 Questions 67 Answers 0 Followers
Questions related from Khan Md Taufiqul Arif
In particular I'll be looking for a panel of miR-SNP, this panel will be used in a case-control association study. I have collected info on chromosome no and location, MAF, ancestral allele, Gene,...
29 September 2017 4,558 3 View
To design a kinase inhibitor which criterion should we follow.. 1. Solitary target with number of compounds 2. Multi-target with single compound Off-target effect might be detrimental but...
30 December 2016 3,353 8 View
The compound we have been using has been showing strong anti-inflammatory activities in dermal fibroblast cell, we believe that it might have shown the similar role in cornial epithelial cell....
08 December 2016 9,231 2 View
For the last few experiments I am getting no elevation in ROS comparing to the control group (No TNF-a). Generally I used to get ~30% increase (Amplex Red Assay. I am keeping the cells starved for...
26 September 2016 4,648 6 View
Measuring intracellular ROS using DCFDA is creating lots of misleading data, I would really like to switch to other protocols available to measure ROS. But my point is where should I really look...
01 September 2016 3,136 3 View
I have been treating HUVEC with my compound to see if it can reduce the TNF-a induced ROS. Usually I treat the compound twice (with DCFDA and after washing with TNF-a). Since the results were not...
05 August 2016 3,497 3 View
To get the best ROS generation (i am expecting 40-50% increase than that of control), how long my HUVEC should be starved and what would be the serum percentage? i am using EBM-2 (Lonza) without...
29 July 2016 2,284 4 View
Previously I tried with EBM-2 (cell: HUVEC), but I found that in cell free condition EBM-2 in presence on DCFDA (20uM) gives very high fluorescence. My plan is to replace EBM-2 with HBSS. But, I...
27 July 2016 8,335 6 View
For some reason I had to use HUVEC with high passage number (p8-p14). I treat TNF-a (10ng/ml) for 1hr and administer my compound to see changes in the ROS level. In P5-P7 I got mostly 25%...
20 July 2016 5,948 2 View
I have been working with a tyrosine kinase inhibitor, which showed significant positive effect by declining ROS generated by TNF-a (10ng/ml, 1hr) in HUVEC. I am using 20uM of DCFDA +plus compound...
15 July 2016 4,795 6 View
Need to detect p47phox in the membrane fraction. But, the protocol I am following giving nothing in WB (lysate, cytosolic fraction and membrane as fraction). Not sure whether the buffers are...
08 July 2016 693 3 View
In vitro, TNF- induces a very weak oxidative response by neutrophils but strongly enhances ROS release on exposure to a secondary stimulus, such as the bacterial peptide fMLF. is it capable of...
04 July 2016 7,148 2 View
Need to separate the membrane fraction for detecting p47phox translocation. I've found that 100,000g for 60min can give satisfactory result. But, the ultracentrifuge we are using now cannot go...
28 June 2016 1,368 1 View
I am culturing HUVEC up to 80% confluency (EGM-2; Lonza), then let them serum starved (EBM-2; Lonza) for 24hr and treated with TNF-a (10ng/ml) for 30min. With DCFDA I didn't get that much of...
17 June 2016 4,451 5 View
Need to know two things 1. What is the appropriate time frame to record RLUs? (i am using a microplate reader, Spectramax M2, can't set the recording time below 1min) 2. How will be the typical...
17 June 2016 6,297 1 View
Trying to measure protein concentration for Lucigenin assay. But even culturing at more than 80% confluency, after centrifugation and sonication getting only 330ug/ml of protein. Hard to maintain...
15 June 2016 1,395 2 View
I am not controlling any of these factors, but feeling a bit shaky. Do I need to maintain a certain wavelength and what would be the optimum temperature for that? Thanks in advance.
07 June 2016 2,877 4 View
The compound is a Tyrosin Kinase Inhibitor (type 1). Can also reduce ROS (in HUVEC, TNF-a induced). I need to know which factors are required to be considered/proved in vitro and in vivo to show...
01 June 2016 7,799 1 View
i have been using lucigenin (5uM; final conc.) preppared in Krebs-Ringer-HEPES (modified) buffer. My stock is 12mM (in acetic acid) and for working I prepared it as 40uM in the same buffer. But...
23 May 2016 3,519 2 View
difference in terms of total ROS and/or type of ROS, after TNF-a challenge
12 May 2016 2,486 5 View
i would like to test my compound in vitro. which cell line should I use to generate atherosclerosis? to prove my compound's (Kinase inhibitor) potency which assays would be rational ?
10 May 2016 1,780 3 View
is there any optimum time frame for that?
09 May 2016 8,427 5 View
Need to find out the specific pathway by which collagen is increasing ROS. What could be the possible technique?
18 April 2016 5,977 4 View
I administered Ang-1 as 50ng/ml for a kinetic assay for 1hr in HUVEC (passage 9) in Collagen-1 (200ug/ml) coated well, but the fluorescence (in arbitrary unit) was very high indeed. Almost 1000 at...
14 April 2016 5,676 1 View
HUVECs are being starved for 24 hr (in 96-well) and same volume to of DCFDA is being added directly. Do I need to replace the media before adding DCFDA? Can cytokines cause any effect on ROS...
11 April 2016 6,350 2 View
I am treating TNF-a to produce ROS in HUVEC. I started with passage no. 5 and used up to passage no. 9. But not sure whether I am getting the same response. Need your suggestions regarding this...
08 April 2016 4,777 3 View
at present I am treating Angiopoietin I to Tie-2 mediated ROS generation for screening my lead compound. But need some more solid ideas about the receptor tyrosine kinases (RTKs) that are involved...
30 March 2016 783 12 View
need to explain to you: TNF-a was prepared as 100ng/ml using EBM-2 (basal) to treat my HUVEC for ROS generation. But the solution was diluted to 10ng/ml and the rest of the volume of 100ng/ml...
22 March 2016 9,311 4 View
I am using 0.05% Trypsin/EDTA. Added 2ml to T75 flask, incubated for 3min but not more than 10% cells were detached and there were very less circular cells. Not sure what was wrong.
16 March 2016 4,906 9 View
My HUVEC is from Lonza. Right now I am using EGM-2, Lonza, but due to its high price now looking for something cheaper. My stock is in EGM-2, can I resuspend them in M-199?
16 March 2016 7,164 4 View
my drug doesn't last long (~1hr at best) in the blood stream, so need know how long the lps actually remains there to cause sepsis. I found that treating my drug (20mg/kg bw; i.p.) after 1hr of...
14 March 2016 4,039 16 View
Assay: Cytokine profile and Survival rate determination Animal Model: C57BL/6 mouse (6weeks) LPS: E. coli: O111:B4, O55:85, O127:B8, Salmonella enterica ser. enteritidis/typhimurium Dose: 15mg/kg...
11 February 2016 733 3 View
for isolating rat aortic smooth muscle. can i use collagenase I?
04 February 2016 3,888 1 View
Need to know the concentration, dosage and preparation of LPS, I am going to test my drug whether it has any anti-sepsis action.
01 February 2016 5,762 3 View
going to start culturing HUVECs using EGM-2 for my experiment and now in need of every detail.
25 January 2016 9,288 7 View
I have found that a tetrameric form of Ang-I would do good but, what is the molecular weight of that particular state and to which company I should look for.
21 January 2016 3,346 2 View
Non-fibril form of Collagen-I
22 December 2015 9,141 2 View
Model: Balb/c mouse (male) Compund: CCl4 Methods: 1. s.c injection: 0.5ml/kg bw, thrice/week upto 4 weeks 2. i.p injection: 0.5ml/kg bw, twice/week upto 4 weeks 3. oral gavage: 0.5ml/kg bw,...
10 December 2015 8,593 10 View
Treatment procedure; formulation, application mode, concentration
22 October 2015 4,929 3 View
I am looking for any biologically relevant oxidants other than H2O2....
21 October 2015 9,014 3 View
I am using both soluble collagen form (20 micro gram/mL, serum free DMEM, over night incubation)and the coating techniques (200micro gram/well, 4 hrs incubation).
19 October 2015 8,323 2 View
UV-B irradiated ROS screening (DCFDA method) in HEK/DDR2 cell line after collagen type I treatment
25 September 2015 8,761 0 View
I have been searching for SNPs in miRNA genes (formulated panel of 65) in a designated breast cancer population by iPLEX/MALDI-TOF-MS. But after that just got stuck into and couldn't go...
01 January 1970 3,746 1 View
Dear Tuan Zea Tan, My research is focused on association of miRNAs to the molecular subtypes of breast cancer and their clinical significance. I'd really appreciate if you could share the...
01 January 1970 3,894 2 View
