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Questions related from Abdul Mannan
I am treating MCF7 cells for 96hrs and my protocol is as follow 1-Day 1- Seed cells @ 2.5x10^3 cells/100ul per well in 96 well plate in 10% FBS containing DMEM complete media 2- Day 2- Drug...
08 August 2017 169 5 View
We are planning to do BT474 xenografts in Nude mice. Though most of the article mentioned of using E2 for BT474 xenografts, I came across some other article in which the use of E2 was not...
05 May 2017 2,876 1 View
I am doing some experiments with the MCF7 breast cancer cells. I have established MCF7 cell lines with and without suppressing the protein of interest. Currently, I am doing cytotoxicity and...
04 April 2017 4,568 4 View
We are working on breast cancer with the main focus on luminal B tumours. We have been using BT474, ZR751 and ZR7530 cell lines as a model for luminal B breast tumour, but recently I came across...
01 January 2017 8,675 2 View
We are trying to do soft agar assay in BT474 (luminal B) breast cancer cells. I tried couple of times, but could not get even a single colony. Last time I even tried using MCF10A-Ras cells but...
03 March 2016 3,881 10 View
I am doing Transwell matrigel Assay on BT474 cell lines as follow To coat trasnwell inserts (8um pore) I added 75 ul of 2mg/ml matrigel in serum free media and incubated in the incubator for about...
12 December 2015 6,911 7 View
We are doing cell cycle analysis in ZR-751 cell lines. We seed cells @ 5x105 cells per well in six well plate on day 1. On day-3 we are treating cells with DNA damaging agent for 1 hour followed...
07 July 2015 6,717 5 View
Hi, I did soft agar assay on MCF-7 cells. I used 0.7% and 0.5% for plate 1 and for plate 2 0.5 and 0.35% of agarose for bottom and upper layers respectively. 5000 cells were seeded in each well....
04 April 2015 8,330 1 View
We want to standard curve for luciferase assay by using a range of 5x106 to 1 cells / well in 96 well plate. We will use Kit from Promega. My question is since upper limit is too high to high to...
04 April 2015 6,556 3 View
Hi, We are trying to do soft agar assay in six well plate and performing on MCF7 breasr cancer cells. We are preparing bottom layer with 1.5 ml 0,8% agarose land top layer with 0.35% agarose. At...
03 March 2015 3,284 7 View
I am doing western blot and the problem that I encounter is that the target protein and housekeeping proteins' molecular weight is close enough to cut the gel to probe on separate membranes....
08 August 2014 3,486 9 View
I have designed a pair of primers for my target gene to determine its expression by Syber-green kit. Both primers are spanning two across exons. However, when I BLAST primer pair in NCBI, one...
06 June 2014 1,759 0 View
I am using K562 cell line. Recently, I tried to transfect these cells with siRNA bought from qiagen (flexitube siRNA). For optimization I used forward transfection method in twelve well plate. We...
05 May 2014 2,614 5 View
I run western blot gel and protein maker look really fine after running. Then I transferred the protein to PVDF membrane in transfer buffer with 20% methanol. But after transfer, protein marker...
03 March 2014 3,317 15 View
Viral mediated gene delivery is a common practice to transduce normal cells into cancerous cells, such as the transfection of hematological stem cells with MSCV-BCR-AB-GFP-IRES vector. My question...
03 March 2014 5,778 0 View
We are planning to treat mouse model of CML with Imatininb. We want to prepare a water base Imatininb stock solution and my question is: How do we sterilize that solution? Should we prepare by...
02 February 2014 1,739 6 View
We are planning to induce chronic myeloid leukemia in mouse. Our plan is to induce CML by infecting bone marrow cells from donor mouse with retroviral vector, and then implanting into recipient...
12 December 2013 7,084 2 View
We are planning to induce chronic myeloid leukemia in mouse model with the MSCV-BCR-ABL-ires-GFP retroviral vector, but I am unable to find any company who is selling complete retroviral vector...
12 December 2013 4,782 2 View
