Why is my GOI not expressing after lentiviral transduction despite RNA increase and protein is detected in transfection?

Hi, I'm trying to create a stable overexpression line with EF1a-FlagGOI-IRES-GFP using lentivirus packaged with pxPAX2 and pM2G. After transduction of 293T with 8ug/ml polybrene, I was able to...

29 January 2021 1,878 4 View

How to correct a deviation in the molar ratio in an Isothermal Titration Calorimetry equipment?

Our equipment of ITC does not reach the molar ratio (N=1) of a titration of EDTA with Ca2+ in MES buffer pH 6. We have cleaned with DCN 90 and HCl pH 2, but the N value is very low, around...

10 November 2020 3,137 3 View

How can I obtain the cohesive energy density and solubility parameter for a crystal using the Materials Studio software?

Hello, I am currently analyzing properties of some minerals such as Bi2O3 by MD using Materials Studio, I need to obtain the cohesive energy density (CED) to obtain the solubility parameter,...

02 November 2020 4,356 1 View

In which solvent should I reconstitute mitoTEMPO?

Dear colleagues, I was wondering which solvent should I use to reconstitute mitoTEMPO, I've seen I could use water or DMSO but I'm not sure about the stability of the compounds in the respective...

18 May 2020 2,882 3 View

Does anyone knows if is there any open source repository of historical aerial photography that includes the north of Chile?

My PhD includes a remote sensing survey of the Atacama Desert using freely available satellite imagery to detect archaeological sites. So far, I have not found any source of historical aerial...

12 May 2020 1,383 8 View

Converting Large Chemical Formats to .pdbqt for AutoDock Vina?

I am trying to convert a large single file library (.sdf, .pdb) containing 50k+ molecules into a .pdbqt file for screening. I have tried using AutoDock Tools but it crashes upon trying to load the...

26 April 2020 5,020 6 View

Do you think the approach to sustainability and environmental issues will change for human society after this pandemic?

We need to be closer to the nature and don't look for money and other values.

29 March 2020 1,238 4 View

Which statistical analysis should I follow for analyzing the effects of two factors in two different cell types cultured in vitro?

Hello all! I have performed experiments involving two different cell types. These two cell types have been cultured in vitro and measurements have been performed after 4, 7 and 10 days of culture....

18 March 2020 6,337 10 View

Can Bacteria grow in DMSO 99%?

I used to test antiproliferative activities of chemical compounds dissolved in DMSO. I have been surprised when testing one of this compounds in HBL-100 cell line bacteria contamination appears. I...

13 February 2020 4,542 2 View

Does anyone know how many samples can I sequence with the minION basic starter pack ?

02 December 2019 3,228 1 View

IHC on mouse liver hepatocytes?

Can anyone help me choose primary antibodies for mouse liver hepatocytes and also in conjunction Cd8 cells. Also Cd3 cells. I want to look at hepatocytes and adjacent immune cells in mice with...

01 March 2021 6,953 1 View

Anyone know of a good progesterone antibody?

I am looking for a good progesterone antibody to quantify levels of progesterone in zebrafish. Applications would be ELISA, and immunohistochemistry.

28 February 2021 5,929 2 View

Reagents for histology slides?

Dear all, I am planning to study histology and want to do that at home. I have only stained a blood smear with Giemsa but now want to try a wider range of animal tissues. What reagents will I...

24 February 2021 5,430 3 View

IF troubleshooting: looking at an intracellular protein that localizes to the cell's and how to reduce increase in signal due to cell ruffles?

I am currently trying to stain for PI3KBeta in MDA-MB-231 cells. My current issue is that 1) beta stains all over the cell and 2) there is an increase in signal at the edge of the cell and this...

24 February 2021 8,582 1 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

What's the difference between 0.1% triton-X100 and ice-cold methanol in the immunofluorescence assay for intracellular protein staining?

Hi, Guys, I noticed that researchers usually permeabilized the cells with 0.1% Triton-X100 or ice-cold methanol (90%) for an intracellular protein staining, it looks like that the two methods work...

15 February 2021 5,606 4 View

Troubleshooting with immunofluoresence/confocal microscope? What went wrong?

Hi guys, I have no experience and little knowledge on immunofluorescence and confocal microscopy. I tried to visualize the expression of FXR in my cells (THP-1 macrophages), yet I did not...

11 February 2021 562 6 View

Does anyone have any experience with the Motic EasyScan Pro Slide Scanner?

I am interested in purchasing a slide scanner for histology and was presented with the Motic EasyScan Pro but I have never heard of the company, nor know of anyone who has used the system. Can...

11 February 2021 2,416 2 View

How to do immunofluorescence staining of the microtome sections of paraffin-embedded tissue ?

I plan to do immunofluorescence staining of microtome sections of paraffin-embedded metastatic tumor tissue with two or more than two antibodies with different emission spectra. Should I follow...

10 February 2021 3,113 3 View

How do I fix tissue rolling while cutting frozen rat brain section with a cryostat?

I am cutting rat brain sections at 20um at -20C. The tissue is cutting nicely out of the O.C.T solution, but I can not get a decent section to adhere to the slide. I have included an image.

09 February 2021 1,345 5 View