What sentence do I have to put in the calculation entry to be able to make corrections for overlapping bases (BSSE)?

computational chemistry: I want to make the base overlap corrections for the dichloroacetyl chloride dimer. I put the following sentence to run the calculation in Gaussian: frec B3LYP/6-311++G**...

27 February 2020 6,347 3 View

What binding buffer would you recommend for NI-NTA superflow column purification?

I have to start the purification of a protein using a NI-NTA superflow column. My protein has an aproximate pI of 7. I have no experience with IMAC purification, i know i may have to try...

07 January 2020 3,884 4 View

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...

02 January 2019 4,794 4 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:...

08 May 2016 6,236 1 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...

03 May 2016 2,054 1 View

What are duck egg patterns in inverse distance interpolation, and why it occurs?

25 April 2016 604 3 View

What makes Runge Kutta Method more accurate than Euler method for solving ODEs?

Runge Kutta method gives a more stable results that euler method for ODEs, and i know that Runge kutta is quite complex in the iterations, encompassing an analysis of 4 slopes to approximate the...

02 April 2016 5,304 18 View

What is the function of the ozone scrubber in some ozone analyzers that use UV absorption?

I understand the process of UV absorption, but I am not sure about the Ozone scrubber, is it just a matter of comparison, like a blank for the final result of ozone concentration in air?

22 January 2016 3,473 4 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...

28 April 2015 754 2 View

Can anyone suggest a protocol for EDS (Ethane dimethane sulphonate) preparation?

04 March 2015 8,537 2 View

Does anyone know an alternative product for cardiac Troponin T antibody for Immunofluorescence (Thermo # MA512960)?

We have been using Thermo Fisher anti-cardiac Troponin T antibody for immunofluorescence staining in FFPE mouse heart sections but it was recently discontinued. The literature shows it to be the...

01 March 2021 4,937 1 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

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

Markers of mitochondrial activity in FFPE tissues?

Hello everybody, Does someone have any experience with assessing/extimating mitochondrial activity on FFPE tissues? Precisely, no TOM20, ATP5B, etc: we are trying to get an approximative idea of...

24 February 2021 4,618 4 View

Can a Golgi-stained dendritic spines picture processed by Image J be directly used for publication?

24 February 2021 3,348 2 View

Is there an appropriate CD8 antibody for IHC-P for staining human breast cancer tissues?

Hello, We are planning on staining Formalin-fixed paraffin breast cancer tissue sections (FFPE) with CD8 Ab via immunofluorescence (Tissue IF). Based on our existing protocol and imaging...

24 February 2021 9,578 4 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