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Questions related from Amanda Rocha
I have seen that some groups use CSFE to stain some parasite species before infect cells to detect percetage of infected cells. Could I use CSFE to stain Trypanosoma cruzi and measure infection in...
16 August 2020 2,233 2 View
I have purchased THP-1 cell line from ATCC and they recommed mercaptoethanol. It is a reducing agent, what is the function of this kind of agent in culture medium of THP-1 cells? How...
29 May 2019 8,308 7 View
Is it best perform fixation step with formaldeihyde or paraformaldeihyde at 4ºC or room temperature or there is no great difference? I work with human mo-DCs in culture and I need to detect...
13 May 2019 3,730 3 View
I need to detect and measure the abundance of cytoplasmatic proteins (AIM2, IFI16, GBP1, GBP4) using direct detection by flow cytometry with fluorochrome-conjugated antibodies in human mo-DCs and...
12 May 2019 1,687 4 View
I need to detect and measure the abundance of cytoplasmatic proteins (AIM2, GBP1, GBP4) and some nucleic proteins (IFI16)using direct detection by flow cytometry with fluorochrome-conjugated...
12 May 2019 9,235 2 View
12 May 2019 8,252 5 View
Which methodology is most recommended to perform a gene knockdown in human monocytes- derived dendritic cells: morpholino or siRNAs?
31 January 2019 4,009 5 View
I want to detect cleaved caspase 1 in human monocyte-derived dendritic cells after infection with my pathogen
15 January 2019 4,704 5 View
Initially, I have thought to use siRNA to study the role of specific genes in mo-DCs infected with T.cruzi, but I found this work these days:Induction of the interferon response by siRNA is cell...
18 December 2018 908 3 View
I have pX330 available in my lab, but I haven't found many recent works using this plasmid.
11 November 2018 9,473 3 View
I have tried to established the amount of RNA input in my RT-qPCRs reactions to amplify interest targets in human monocytes-derived dendritic cells and I perform a RT-qPCR with 4 different RNA...
20 June 2018 3,705 1 View
These 2 analyze methods are recommended for specific gene expression analysis?
01 June 2018 6,623 4 View
I have analyzed mRNA human samples in bioanalyzer and the RNA concentration provided did not match with the RNA concentration of the same sample in the same dilution provided by Qubit. What RNA...
04 May 2018 2,895 3 View
I am trying to define a standard curve for the primers that I have designed for qPCR and I ordered to sythetize the correpondent oligo for test the primers in different concentrations, but I don't...
29 November 2017 5,400 2 View
I need to determine infection rate of dedritic cells by Trypanosoma cruzi determining amastigotes forms insides these cells and I have thought to do that by RT-qPCR to detect some specific...
20 May 2017 7,879 4 View
Dendritic cells can produce IL-4?
20 May 2017 4,170 2 View
I have gotten a low RNA amount from human dendritic cells using RNeasy kit from Qiagen, such as 11ng/ul and I need to perform a qPCR assay from these samples. Or trizol still is the better way to...
22 February 2017 1,062 3 View
I have tried to isolation RNA from human dendritic cells using RNeasy kit (QIAGEN), but I got a very low yield of RNA, such 30ng/ul. In the last time, I tried to combine QIAGEN protocol with...
19 January 2017 8,448 3 View
I would like to know what cytokines are produced by dendritic cells. I need to know ,specifically: These cells can produce IL-1beta, IL-6, IL-10, IL-12p40, TGF-beta, and TNF-alfa?
15 January 2017 3,238 3 View
Currently, I use the protocol of Ficoll in order to obtain PBMCs from the whole blood colected from the donors. I would to know if there are alternative ways to isolate PBMCs.Percoll have the...
12 January 2017 4,013 3 View
In the lab where I work, it's used the experimental protocol of Canavaci et al (2010) in order to elicit metacylogenesis in epimastigotes from CL Brener strain cultivated in LIT medium. Then,...
11 January 2017 3,274 2 View
I use beads conjugated with anti-CD14 in PBMCs samples in order to purify monocytes through a column linked to a magnetic board, which is obtained from MACS. However, using this kind of kit is too...
11 January 2017 8,169 5 View
I have performed a antibody titration to detect a specific cytoplasmatic protein in human monocytes starting with the double amount of general antibody recommendation (2 ug) and I have made 9...
01 January 1970 752 1 View
I have observed that both index are used in order to evaluate the best concentration of antibody in FC, but which one is most recommended to discover the best antibody concentration that provide...
01 January 1970 2,886 4 View
I intend to silence a gene in human monocytes-derived DCs or THP-1 cells using RNAi to study its role in a infection. Where I can purchase validated and reliable siRNAs to silence a target gene?
01 January 1970 9,063 2 View
I have found 2 protocols for caspase 1 detection from cells supernatant and both said to avoid fetal calf serum (FSC) or other kinds of serum to the media before the stimulus and to collect the...
01 January 1970 4,840 2 View
