Normalization by DNA content in glucose stimulated insulin secretion assay for islets?

I want to ask about normalization step: if I want to normalize by DNA content how I should isolate DNA after treatment of the islets with acidified ethanol?

07 August 2017 4,407 0 View

Has anybody faced difficulties with islets isolation from db/db mouse model?

Recently, I am trying to isolate islets from db/db mice. I use the same protocol I regularly use for islets isolation from mice but I can't get any islets or very few pale islets attached to...

01 February 2017 5,872 7 View

For murine islets, can I work on islets extracted from one mouse or I have to pool from several mice?

I am recently extracting murine islets. I see most researchers recommend pooling islets from several mice. I just want to check expression of my protein of interest in islets as an initial stage...

10 November 2016 9,330 3 View

What is the best internal control for western blots of mice lysates?

09 October 2016 6,228 3 View

Can I treat mice islets expressing tamoxifen induced cre in vitro with tamoxifen to knock out a floxed gene?

I have a mouse model that expresses modified Cre recombinase, so that only upon tamoxifen treatment of the animals they express Cre, thus KO of floxed gene can be induced. I want to isolate the...

07 August 2016 2,612 2 View

I am asking if somebody previously transfected islets of Langerhans isolated from mice with siRNA before. Is this going to work?

If this works what kind of tranfection reagent is recommended for the islets?

07 August 2016 4,448 9 View

Are beta actin and tubulin levels in islets affected by fasting ?

I am interested in knowing the expression of a certain protein in the islets under feeding and fasting conditions. I use the same total protein concentration for immunolotting. In my first trial I...

07 August 2016 4,264 1 View

I want to ask about measurement of insulin content in MIN6 cells?

I did a glucose stimulated insulin secretion assay for MIN6 cells. I measured the released insulin in the Kreb's Ringer buffer containing certain concentrations of glucose. In the end I collected...

06 July 2016 3,844 2 View

Which transfection reagent is best with reporter assays on MIN6 cells?

I want to transfect PGL3 plasmid to MIN6 cells. I tried Trans-it trnsfection reagent but didn't work well.

01 January 1970 5,237 2 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

No title

02 March 2021 3,060 3 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

IHC on mouse liver hepatocytes?

Can anyone help me choose primary antibodies for mouse liver hepatocytes and also in conjunction Cd8 cells. Also Cd3 cells. I want to look at hepatocytes and adjacent immune cells in mice with...

01 March 2021 6,953 1 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

How to avoid ulcers in s.c. injected tumors ?

Recently, we have injected s.c. B16F10 tumor cells in BL/6 mice. At around 12-15th day some animals showed ulcerations in tumors , with a big hole in the middle (it looks like a volcano). We have...

01 March 2021 9,938 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

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