Which one is the better plan for time course study?

I am doing a time course experiment. Treat cells with a drug for different times (0h, 2h, 8h, 18h, and 24h, respectively), and then harvest cells for RNA to check cell responses. So here we have 2...

31 December 2016 4,533 9 View

What any effects of stearic acid treatment on protein concentration?

02 March 2016 4,914 3 View

Do you add M-CSF or L929-conditioned medium to cell culture when you treat BMDM cells?

02 March 2016 9,974 11 View

Does anybody have good idea with three plasmids co-transfection into macrophage cells?

I am trying to co-transfect 3 plasmids into macrophage cell line (RAW264.7). I can transfect single plasmid successfully and can detect with immunoblotting. But it is very hard for me to do...

09 October 2015 5,465 8 View

What 2x Laemmli Sample Buffer recipe is better?

Up till now, there are two kinds of 2x Laemmli sample buffers: Buffer 1) 65.8 mM Tris-HCl, pH 6.8, 2.1% SDS, 26.3% (w/v) glycerol, 0.01% bromophenol blue Please refer to the original...

04 May 2015 1,878 8 View

How do you make acetylation detection easier with Western?

I treated macrophage cells with LPS (1h to 24h time course) and tried to detect p65 acetylation. Method 1) I collected total cell lysate with RIPA buffer containing 10mM NAM and 10uM TSA. And then...

31 December 2014 7,371 8 View

What is the best method or kit to purify mono-antibody from tissue culture?

08 September 2014 3,162 4 View

Can anybody recommend antibodies against Human E-Selectin, VCAM-1 and ICAM-1?

I am working with HUVEC cells and hope to detect E-Selectin, VCAM-1 and ICAM-1 protein expression with western blot and cell ELISA (for surface expression). Does anybody have experiences with...

04 May 2014 2,692 4 View

Can anybody help me with western blot using small intestine tissue lysate?

Here is my protocol: 1. Collect small intestine from mice and push out the stool. 2. Flash freeze in liquid nitrogen and then store the tissue at -80C for later use. 3. Homogenize tissues in RIPA...

02 March 2014 4,307 9 View

Does a forest free freezer really destroy antibodies?

All freezers (-20C) we have are forest free. Is it safe to keep antibodies in those freezers? If safe, how many months?

02 March 2014 426 9 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Benefits of the 24 welled plate?

We have always used the 96 welled plate, the 48 welled plate but apparently there is much evidence on the 24 welled plate and above all its benefits.It states it is the same you would use on ELISA...

28 February 2021 1,149 1 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Can two EC50 values with different hill slopes be compared?

Hello, I fit a classical dose response curve Y=Bottom + (Top-Bottom)/(1+10^((LogEC-X)*HillSlope)) to my data (not related to inhibitors etc). The model I use is purely qualitative but it fits...

23 February 2021 8,155 5 View

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...

17 February 2021 4,844 4 View

What is the best temperature to coat an ELISA plate?

Hello, I want to control the temperature of my ELISA plate during incubations to get consistent results. In general, what is the best temperature to coat the plate with antibody? I have been...

16 February 2021 2,377 7 View

How to visualize stress in a coating defined by "Skin" in Abaqus?

Hello All! I'm trying to model a contact between a coated substrate and a tip in Abaqus. The coating is ultra-thin compared to the geometry of the substrate. So I defined the coating using the...

16 February 2021 1,594 3 View

2D wavelet transform?

Dear all I have a 2-dimensional mode shape of one concrete beam and I put the data of mode shape in a matrix (69*3) which the first column in the matrix is x coordination of my beam and the second...

10 February 2021 3,301 3 View

Should the AUC in a concentration-time graph of a subcutaneous implant be roughly the same for poeple?

I have 2 concentration time curves of a subcut drug implant. the first group has a peak around the 50 day mark where as the second group does not. Both groups received the same implant by the same...

10 February 2021 2,305 3 View

Is there a way to tell based on appearance which protein crystal should diffract the best?

I have had unheard of success with protein crystallography lately from a super successful protein expression and purification batch. I have attained a lately reproducible vast amount of crystals...

09 February 2021 5,155 5 View