Could anyone tell me the best parameters for transfer of a 7kDa protein? Nitrocellulose vs PVDFat 2uM?

09 October 2014 508 3 View

What fraction of proteins within a cell are of low molecular weight (<15kDa)?

09 October 2014 1,614 4 View

Does someone know if Ponceau S will stain low molecular weight proteins (~7kD)?

Unable to see small proteins on my transfer to PVDF membranes.

08 September 2014 2,198 4 View

Will Ponceau S bind proteins of low molecular weight, i.e. less than 10kD?

Ponceau S staining

08 September 2014 6,538 8 View

What fraction of C6/36, or any cell line in general, consists of low molecular weight proteins (less than 10kDa)?

With Ponceau S staining I have not, as of yet been able to see any bands below 10kDa (green band) on a .45um PVDF membrane after 45, 30, or 15 minutes of transfer at 100V and 400mA using a 15%...

08 September 2014 9,390 0 View

Does anyone know which would promote better transfer for low MW proteins (~7kD)?

I'm trying to transfer a low molecular weight protein from a 15% SDS-PAGE gel to a 0.45um PVDF membrane. I've heard many people suggest using a 0.20 um membrane, but many others suggest that it is...

08 September 2014 237 5 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

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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

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

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

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28 February 2021 9,936 2 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View