Why do PC12 cells die after NGF addition?

I have been working with PC12 cells for years. Lately, however, I noticed that they start dying after 3 days of differentiation with NGF . My protocol is the following: I plate the cells onto poly...

31 December 2018 5,078 1 View

How much time to treat PC12 cells with KT5720?

I am using KT5720, a PKA inhibitor. The first time I used it, it had no effect at 1 uM 1 hour before my experimental treatment in PC12 cells. I would like to know what is the optimal time of...

08 September 2017 1,656 1 View

How can I establish the optimal primer concentration for qPCR with SYBR Green?

03 April 2017 3,463 8 View

What is the best endogenous control for differentiating PC12 cells?

I am working with PC12 cells and I take mRNA samples every 48 hours after differentiation with NGF. Which would be the best endogenous control for qPCR given that it needs to be one that does not...

04 May 2016 4,634 1 View

What are the best conditions for Western Blotting alpha synuclein?

I am working with PC12 cells and would like to know what are the best conditions to perform western blot of alpha synuclein (14 kDa). Especially, regarding: best buffer for protein extraction:...

04 May 2016 8,212 1 View

How should I perform anti alpha-synuclein western blots?

I am working with PC12 cells and. My problem is that when I perform anti alpha-synuclein Western Blots, the signals of this protein are never consistent with each other, even when I run duplicates...

05 June 2015 771 5 View

What is/are the best agent/s and protocol to differentiate SH-SY5Y cells into dopaminergic neurons?

I want to test effects of a toxic drug on SH-SY5Y cells and I read that there are many agents that induce differentiation into different phenotypes but I could not conclude which of them are...

03 April 2015 5,126 3 View

Can you help me figure out my problem with alpha synuclein Western Blot?

I am trying to perform western blots of alpha synuclein (14 kDa), using wet transference 1h 15min at 85 V, but I get this weird "patchy" pattern, the band isn't a whole defined band. What could be...

03 April 2014 1,926 15 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...

03 March 2021 5,978 5 View

No title

02 March 2021 3,060 3 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