16 Questions 27 Answers 0 Followers
Questions related from Abdullah Al Tarique
I digest my plasmid with KpnI-NotI. I cut the expected 4kb band from the gel, purified it and then I did ligation [1:3 and 1:5 vector:insert ratio] and transformation. I found 18 transformed...
01 July 2024 4,006 0 View
My vector size is 5.1kb and insert size is 4kb. Initially, I performed digestion of the insert from a different plasmid and the vector using KpnI-NotI. I checked them on the gel, excise and...
20 May 2024 3,211 8 View
My vector size 5.1kb and insert size is 4kb. I performed double digestion of vector and insert, ran them on the gel and then extracted them from the gel. 260/280 ratio was good. Then I did...
07 May 2024 8,331 11 View
While preparing E. coli DH5alpha competent cells, most of the methods describe using E. coli DH5a at OD 0.4-0.5. Is there any specific reason for using OD 0.4-0.5? If OD is more than 1, will it be...
18 April 2024 2,318 3 View
I am interested to measure mitoROS using mitoSOX. Lots of paper measured mitoROS using microscopy. Is it possible to measure mitoROS using plate read?
04 May 2023 8,981 2 View
I am looking for antibody against human cell membrane which will circularly stain the cell membrane only. Could anyone name such antibody? I want to avoid F-actin, microtubulin bcz they stain all...
23 January 2018 5,240 11 View
I am using phrodo E. coli bioparticle for phagocytosis. The assay is carried out at 37C. I am wandering how to stop phagocytosis activity of cells. I tried fixing cells with 4% PFA and BD-CytoFix...
01 June 2015 1,679 7 View
I am currently struggling with cell fixing and followed by surface staining. Due to my experimental setup, I had no other choice to fix the cells first and then perform surface staining. I used 2...
26 May 2015 1,070 6 View
I fixed my cells with 3.75% PFA at RT for 10min, washed them with FACS buffer, blocked them and finally stained them with CD80, CD64 and CD11b. I found CD80 expression is down compared to unfixed...
15 May 2015 4,025 4 View
And when BM-macrophages or PM are chosen for study?
13 November 2014 5,975 6 View
Is it possible to separate human M1 and M2 macrophages by flow cytometry? Has anyone done this? Could anyone provide any related literature?
10 November 2013 4,987 14 View
I am wondering about the optimum temperature for cell staining with fluorochrome tagged antibody for Flow cytometry. Majority of the articles report staining at 4C or on ice, however, some...
29 August 2013 8,199 5 View
I am doing flow cytometry using human macrophages. I was wondering, which way is more appropriate to place a gate in FSC-SSC plot? Is tight gate better than wide gate? See the attachment.
18 June 2013 5,028 19 View
Lots of murine M1 and M2 macrophage markers have been described in literature. However, the same markers are not translated in human. The markers described for human M2 macrophages include CD163,...
16 May 2013 5,624 9 View
I am planning to isolate monocytes from blood using MACS. I was wondering whether positive selection could have effects on subsequent experiments. Previously I used Miltenyi CD14+ MACS beads. Does...
04 December 2012 4,585 6 View
I am planing to differentiate human monocytes into macrophages by M-CSF. While doing flow cytometry, I want to exclude DCs from macrophages to get a pure macrophage population. I am wandering...
28 November 2012 6,713 2 View
