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

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 the best way to determine the effect size?

How can you use preliminary data and/or previous research to determine your effect size so that you can estimate power and sample size required for future animal studies/clinical studies?

11 December 2015 3,300 3 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

Resistor definition in Piezoelectric bimorph with ANSYS workbench

Hello, good day! I am trying to simulate similar bimorph piezoelectric harvester with below commands. However, the output power doesn't change with resistor value. Please help to find the reason. ET,10,CIRCU94,0 !SET UP RESISTOR R,1,100000 !RESISTANCE VALUE TYPE,10 !SET ELEMENT TYPE E,1,2 !CREATE RESISTOR BETWEEN NODES 1 AND 2 Thanks in advance Regards,

02 March 2021 0 0 View

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02 March 2021 3,060 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

How to define tie constraint between rigid body and another part in dynamic explicit model of Abaqus?

Hi dear researchers I connected a rigid body into the part of my structure by tie constraint and then I added displacement to rigid body, in static general step every thing is fine and these two...

28 February 2021 3,442 2 View