20 Questions 17 Answers 0 Followers
Questions related from Ahmed Al-Omar
Our purpose it to look for different types of proteins on the membrane, and we have different primary antibodies from different hosts (mouse, rabbit, and goat). In general, is it fine to mix...
22 January 2023 4,008 3 View
We are doing western blot and I want to make sure which protocol is the best according to the protein transfer and blotting step. The normal way is to block the membrane and incubate the primary...
16 January 2023 7,059 8 View
In general when I seed adherent cells to a 6-well plate in density 2.5x10^5 cell/well. The cells after 24h gather together near the borders of the well and a few numbers of cells remain in the...
17 August 2022 1,354 0 View
We analyzed gene expression of A549 cell lines that were cultured in different conditions. The fold change data results of B-Actin as a housekeeping gene is significantly different from fold...
11 August 2022 1,548 5 View
After PBMC isolation and washing, I count the cells of PBMC. Most of the time RBCs exist in the pellet and when I count the total PBMC I am not sure if the counted cells were RBCs or lymphocytes....
16 June 2022 5,425 5 View
I got the EasySep monocyte isolation kit negative selection. After PBMC isolation the number of PBMC cells was about 3x10^7 cell/ml. What is the average number of monocytes that I am supposed to...
09 May 2022 3,367 2 View
Can we put the prepared medium containing stimulatory cytokines like MCSF or interleukins at +4C and re-use it again after few days or we should prepare new medium every time?
09 May 2022 7,174 3 View
In most of the research papers, I noticed that for macrophage polarization of PBMC derived monocytes, the heat-inactivated serum is used during the polarization to M1 or M2. What if I used...
30 January 2022 5,674 4 View
We have Vinorelbine tartrate drug with a 10 mg/ml concentration. How can we dilute it and prepare 1 ml of 2 nM (nano molarity) of drug concentration? The molecular weight of the drug is 1079.1.
09 January 2022 6,088 1 View
I am looking for M1 macrophage marker that is unique and be expressed only by M1 and not by M0 nor M2. the research papers show different ones like CD80, CD86, CD64, and CD40. The monocytes are...
05 January 2022 3,069 4 View
I collected PBMC and cultured the cells in full medium with 10 ng/ml PMA in order to polarize monocytes to macrophages without isolating the monocytes from PBMC. After 24h I washed and discard...
23 December 2021 3,542 8 View
In order to get M2 macrophages, I used THP-1 with 100 ng/ml PMA for 48h, then the cells were rested for 24h in a fresh medium without PMA. Then I put 20 ng/ml IL-4 for 48h. However, after staining...
27 November 2021 2,853 13 View
I will perform flow cytometry analysis for M2 and M0 macrophages. The staining process will be performed at another far institution the next day. So according to storing the tubes. Can I collect...
02 November 2021 2,758 5 View
According to the washing solution buffer that is used in cell staining of the flow cytometer. it should be (PBS+5% heat-inactivated FBS +0.1%NaN3). Can I use PBS + 5% non-heat inactivated serum...
01 November 2021 4,641 3 View
I want to polarize THP-1 to M0, then to M2. After adding PMA and getting macrophages, I need to get the highest M2 percentage (90%) because I can not isolate M2 from other subtypes. For this...
07 October 2021 734 8 View
I have 10 ug of IL-4 stock, and I want to dilute and aliquot this stock. what are the solution that I had to use to dilute and aliquot IL-4 stock? PBS, Distilled water, or other?
07 October 2021 556 1 View
Please I want to make sure about the polarization of THP-1 to Macrophages by using PMA, then the polarization of these macrophages to M2 by IL-4 or to M1 by LPS and INF gamma, does the medium that...
03 October 2021 7,721 2 View
I have suspicious of contamination in my cell culture flasks but I am not sure. at the same time, I need to check my medium whether is contaminated with bacteria or not. so what is the most...
29 September 2021 1,917 2 View
How much volume of 100X (Penicillin 10,000 U/mL, streptomycin 10,000 μg/mL) stock should be added to a total 556 ml RPMI medium (500 ml RPMI + 56 ml FBS) to get medium with 1% antibiotic like most...
11 September 2021 769 2 View
I am doing cell culture of THP-1 cell line, at the first culturing and before incubation, 90% of the cells were dead because they were not stored in liquid nitrogen but in -80 C. How can I...
06 September 2021 1,553 4 View
