18 Questions 9 Answers 0 Followers
Questions related from Eduardo Alvarez
Hello, I hope you are all doing well. I'm recently doing MTT assays to find the IC50 of my inhibitory drug. I have seen protocols that formazan crystals begins to form at 2 hours to 6 hours...
08 November 2021 4,654 3 View
I am very new to quantitative PCR and need some help for this technique. For my experiment, I need to perform a quantitative reverse transcription PCR. I extract the RNA from my pellet and...
06 February 2021 10,041 4 View
Hello, I hope you are all doing well. I have a question regarding my treated cells group. I plan to do western blot and qRT-PCR for my treated cells. However, to save time, I would like to...
07 December 2020 1,612 5 View
Good day, I hope you are all doing well. I was designing primers for Caspase 9 and sometimes, the designed primers in NCBI have transcript variants for the same gene (Caspase 9) but at the same...
06 October 2020 549 3 View
Good day, I hope you are all well. I am new in designing primers for quantitative reverse transcription PCR. I am using NCBI to design my primers and my question is: Do I have to design my...
05 October 2020 7,850 7 View
Good day, I hope you are all doing well. I am very new to qRT-PCR and it is one of the techniques that I must do for my research. So far, from what I have been reading and gathering information,...
05 September 2020 4,283 3 View
Good day, I have seen that our laboratory has a lot of stock of Precision plus unstained standards and very few of Precision plus stained standards. Since I need to run a lot of western blots in...
10 July 2019 2,679 2 View
I have been doing some troubleshooting relating to western blot with the help of a friend. We decided to test if our anti-rabbit conjugated HRP secondary antibodies are working properly by...
29 June 2019 1,504 2 View
I recently performed western blot and blocked my PVDF membrane with 5% non-fat milk. After blocking, I did 3 washes with 1X TBST for around 5 minutes each. Then I diluted my primary monoclonal...
10 April 2019 6,858 3 View
Hello, I have been having a difficult time with western blot lately and I was wondering, what blocking buffer and antibody buffer should I use when using antibodies of Santa Cruz Biotechnologies?...
06 April 2019 2,266 4 View
Good day, I prepare Laemmli buffer 2X from Bio-Rad by adding 950uL of Laemmli buffer + 50uL of B-mercaptoethanol for a final volume of 1mL for SDS-PAGE. Since a lot of Laemmli buffer still...
06 April 2019 3,681 4 View
Before running the SDS-PAGE, I added pre-stained Kaleidoscope on the well corresponding to the ladder, 30ug of proteins from my cell lysate samples and then as another marker, I added again...
02 April 2019 2,032 8 View
Good day, I need to perform a viral load analysis to quantify the number of viral particles/mL found in my samples. The virus that I will be working is with the Human immunodeficiency virus type...
21 August 2018 2,571 4 View
I work with suspension cells, specifically, THP-1. I counted the cells before centrifugation ( directly from the flask) and after centrifugation (pellet). Before I had 10 million cells and after I...
09 August 2018 5,068 6 View
Good day, I have been having problems with cell counting with THP-1 cells. I counted my cells directly from the flask by resuspending the culture media to promote equal distribution of cells...
08 August 2018 9,235 2 View
I am recently beginning to understand more or less how to culture suspension cells, but I get confused in regards with passage number in these cells. My question regards to: Thawing: Whenever...
22 July 2018 2,137 4 View
Good day, I have been recommended to expand my suspension cells, specifically, the THP-1 cells by seeding 5 million cells in 100mL RPMI 1640 media and wait for around 1 week and a half until cell...
20 July 2018 529 2 View
Good day, I have to perform a viral load analysis for some samples for HIV-1. I know how to make a viral stock of HIV-1; however I have no experience in viral load analysis, which i hope i can...
19 May 2018 632 2 View
