I am trying to stably knock out a gene in SY5Y cells using the CRISPR/Casp9 system. After selection with puromycin 99.5% of the cells are dead. Why?

02 March 2016 4,485 13 View

Is it possible to dissolve and store lipids in DMSO alone?

I read papers where they dissolve lipids in Ethanol or in Ethanol/DMSO (1:1, v/v). Is it also possible to store glycosphingolipids in DMSO alone to avoid possible ethanol evaporation?

11 December 2014 7,514 10 View

High background of secondary antibody. Any advice?

I want to perform co-localization studies. My secondary antibody anti-rabbit Cy2 has a very high background. Actually, there is no difference between only secondary antibody negative control and...

31 December 2013 2,323 2 View

Antibody reaction?

In my opinion it is not feasible to use mouse anti-human antibodies for mouse cells or mouse tissue. But I am not an expert in immunology and I saw publications where people used mouse anti-human...

04 May 2013 1,654 1 View

mTORC1 activation

How do people usually check if mTORC1 is activated? By checking if S6K1 is phosphorylated? How does mTORC1 get activated? Also by phosphorylation or is it transcriptionally regulated?

11 December 2012 8,187 3 View

How to solve a problem concerning uPAR KO mice breeding?

Does anybody know anything which can be done to get them back to breeding? Are they too old or could there be another reason as to why they are not breeding?

10 November 2012 8,607 10 View

Detecting sr-b1 in liver cells, Huh7

We have major problems detecting sr-b1 in Huh7 cells (liver cells) using the antibody from NB400-104 from Novus Biologicals. Does anybody have experience with this antibody concerning lysis of the...

03 April 2012 6,728 7 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

How to repeat an operation on an atom selection for multiple times in PyMol?

Hi, I am trying to construct a multi-layer fibril structure from a single layer in PyMol by translating the layer along the fibril axis. For now, I am able to use the Translate command in PyMol...

02 March 2021 4,569 4 View

What analysis to choose with a large neste dataset and clearly skewed or kurtotic distributions?

Hello! I have a dataset of n=3000 nested within 8 countries with approximately 200 or 400 responses in each country. I originally planned to perform multilevel modelling with 4 dependent variables...

02 March 2021 6,865 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

How to avoid ulcers in s.c. injected tumors ?

Recently, we have injected s.c. B16F10 tumor cells in BL/6 mice. At around 12-15th day some animals showed ulcerations in tumors , with a big hole in the middle (it looks like a volcano). We have...

01 March 2021 9,938 3 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

Good morning research fellows, Has anyone worked with senna alata plant extract on any kind of fish?

Literatures on the piscicidal effect of sennaa alata is scanty. if you have any related litrature either in the bark, leaves or roots used as treatment to any aquatic organism please kindly help...

28 February 2021 9,138 2 View

How to plot the probability paper of Gumbel distribution using R programing?

In R-studio, there are many commands of Gumbel package. Arguments are also different. I`m asking about the alpha parameter of the Copula which must be greater than 1. If this is the one used to...

25 February 2021 5,229 3 View

Looking for genomic DNA extraction kits from mouse tissue (centrifuge-based, not spin-column)?

Hello. I am looking to extract genomic DNA from mouse brain and lungs and I need a kit that is not spin-column based because I need to scale up the solution amounts for 400mg tissue (most cap at...

25 February 2021 155 2 View

What are the pros and cons for mice versus rats?

Hi Everyone, I am designing a pilot study using rodents to assess the effects of simulated space environments (radiation + altered gravity) on microbiome and immune system. I have limited...

24 February 2021 1,825 3 View