How do I culture more CD11c+ BMDC from mice?

Hello, I am new to harvesting BMDC and my most recent experiment had 45% CD11c+ cells that were viable (also many dead cells, only 20-30% viable). I followed the method from Madaan A, et al....

06 July 2018 319 3 View

How many times can you strip a western blot?

Hello, How reliable are western blots after they have been stripped? Does the buffer strip all lanes equally? If you load 20 ug onto a gel and strip twice, how much would be left? I'm trying to...

01 February 2018 4,840 3 View

How much EDTA should be used for flow cytometry purposes?

Hello, I've been perusing the literature and a variety of protocols use EDTA concentrations from 0.03-5 mM EDTA in their buffers for isolating immune cells from the lung and/or BALF for flow...

31 December 2017 2,134 3 View

Why does the type of collagenase matter for lung digestion?

Hello! I am attempting to analyze cell populations in mouse lung homogenate using flow cytometry. I have seen a variety of protocols that use a range of collagenase and DNAse concentrations,...

09 October 2017 4,684 4 View

When propagating laboratory strains of Influenza, is ultracentrifugation required for purification?

07 August 2016 8,045 0 View

How do you use cell lysates for studying enzyme kinetics?

Hello, I'm trying to determined the Vmax, Km, etc...for a membrane bound short chain dehydrogenase. I am planning on using cell lysates as a source of the enzyme. Do you know of any protocol to...

03 April 2016 1,925 5 View

What is a solid method to separate nuclear and cytoplasmic proteins from lung tissue?

I've been using commercial kits to separate cytoplasmic and nuclear proteins. I noticed that many advise against using their kits for frozen tissue because the nuclear membrane can break and leak...

06 July 2015 9,735 1 View

How does reaction time effect ligation?

Typical ligation protocols (for inserting a certain gene/deletion into a plasmid) recommend a ligation of 1-24 hrs. Has anyone seen significant differences in yield between 1 hr vs. 24 hr? Thanks

08 September 2014 5,635 3 View

Can anyone help with disappearing plasmids?

I want to know if anyone has ran into this problem before: I'm making a deletion construct and I have positive results in colony screening. When I miniprep the plasmid after overnight culture, the...

07 August 2014 4,079 8 View

I got 02 moderator values defining Johnson-Neyman significance regions. Is it an error?

Hi everyone, I am conducting research for my Master's thesis. I am using PROCESS by Johnson-Neyman to analyze my Moderator model. I test the relationship between Public Service Motivation and...

03 March 2021 2,350 2 View

How to code open-ended questions in SPSS?

Hello all, In SPSS I am going to code 2 open-ended questions. I have already read all the answers and I made a list of the most important categories to which I can code the answers. This question...

02 March 2021 1,757 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

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 View

No title

01 March 2021 3,905 5 View

What test should I use to compare two or three means, if data is not normally distributed but sample size is > 30 in each group?

I personally feel this as a most confusing question. As per my understanding, if sample size is > 30, and sample is skewed, we can still use paramteric approach, as it will follow Z...

01 March 2021 1,587 6 View

Is it necessary to report effect size (ES) in statistics of biomedical research given that we already report p values routinely?

Pls share your thoughts

01 March 2021 7,330 4 View

Is there a method of determining feed digestibility without the need to re-pellet the feed?

I am aiming to determine the ADC values of a commercial fish feed, but I do not have a steam pelleter or extruder available to me. Ideally, the pellets could be coated with some inert marker....

28 February 2021 7,967 3 View

How to eliminate Steady state error from Reinforcement learning genetic algorithm code?

The following code (see 1st 2 images attached) is used to produce PID controller values that are designed to control the system (G). The code finds the PID controller values (noted as k) by using...

28 February 2021 6,560 14 View

Can Ethovision store the generated data and video recordings directly to NAS system?

The size of data generated by Ethovision is huge. Having NAS system for data storage seems to be a good option, I am wondering whether Ethovision can directly store the data to the NAS system...

27 February 2021 6,350 2 View