What are the best practices for live cell imaging?

01 February 2015 1,100 4 View

What are these problems after replacing the Hg Lamp in the epifluorescent microscope?

I recently changed the Hg Lamp in our epifluorescent microscope and now for some reason the CFP and YFP channels do not work, they seem over-exposed even at exposure times as short as 10ms....

06 July 2014 3,565 6 View

Can someone advise on Affinity vs. Dynamic Affinity?

I have been reading some papers regarding the dynamics of receptor-ligand binding for my project and have come across this term "dynamic affinity" quite often and I am wondering if it actually has...

09 October 2013 1,757 2 View

Reagents for transfection of Human Pulmonary Microvascular Endothelial Cells?

I am trying to transfect HPMEC using Lipofectamine 2000. I already tried transfecting for 6h using serum-free media (otimem) and the cell viability dramatically decreased, I reduced the...

08 September 2013 1,406 4 View

How to quantitatively measure intracellular protein concentration?

Does anyone know an accurate technique to quantify intracellular protein concentration besides ELISA? I need a reliable and highly quantitative method to measure protein concentration to gather...

05 June 2013 8,858 4 View

Any suggestions for establishing a Tetracycline inducible mammalian cell line using the TRex system from Invitrogen?

For the last couple of months I have been trying to establish an inducible HaCaT cell line using the TRex system from Invitrogen. I am following the standard protocol (first transfect with...

31 December 2012 3,283 16 View

Are MDA-MB-231 cells sensitive to Olaparib?

Hey!!! Does anybody know whether MDA-MB-231 cells are sensitive to Olaparib? If not, does someone know which cell line is sensitive to Olaparib and what should be the concentration of Olaparib to...

01 February 2012 4,694 2 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Have someone successfully used the ER alpha (Santa Cruz sc-8002) antibody in immunostaining ?

Santa Cruz now recommends this new antibody (F-10) but I can't find a publication using this reference. I'm working on human et non human primate hypothalamus and so far, it doesn't work. Is...

02 March 2021 10,003 2 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

What is the characteristic length to determine the Reynolds number for the scattered turbulent fluid flow?

Dear all, I just saw this example in FB on the difference between laminar flow and turbulent flow. The flow from a pipe hits the sink and scattered. The flow from the pipe is characterized as...

02 March 2021 1,999 5 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

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

How to simultaneously perform multiple biochemical assays?

I am a research scholar working on heavy metal stress on plants. I will be doing biochemical characterisation( protein, carbohydrate, proline, antioxidant enzymes and many more assays) at interval...

01 March 2021 6,999 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

How does an antibody have full intact antibody + half antibody?

Hello, I was just wondering how does a antibody have full intact antibody + half antibody? Is it because of the drug conjugation?

28 February 2021 1,577 1 View