Semi Dry vs Wet Transfer recipe?

Hi I found that there are two different recipes for transfer depending on the mode (Semi-dry or wet) of the transfer. Why is it so? If anybody has prior experience with the use of both the...

11 December 2019 3,859 3 View

Transfer buffer recipe?

Hello I was performing the semi-dry transfer of my gel to the membrane for western blot What is the transfer buffer recipe that you guys use for semi-dry transfer? I saw a different recipe for...

10 November 2019 8,508 3 View

Duration of transfection to induce p53 silencing?

Hi All How long does it take to silence the p53 gene in breast cancer cells? Does the WT or mutant p53 status affect the transfection efficiency? Any other inforrmation regarding transfection...

06 July 2019 752 3 View

Molecular weight of protein is low but signal is at higher molecualr weight ?

The mol wt of my protein of interest is about 27 Kda but i am getting the signal at approx 50Kda. I am Using polyclonal antibody and the standard 12% gel for the detection. Running gel conditions...

05 June 2019 3,753 6 View

Cells found contaminated upon thawing?

Hi I froze the HEK 293 cells some 3 months ago in the Synth-a-freeze medium. When i started thawing them back they are heavily contaminated with bacterial growth. i have lost 4 aliquotes now due...

02 March 2019 8,264 1 View

Ponceau S Stain prior to probing the membrane with antibody?

Hi everyone If i stain my NC membrane with Ponceau Stain prior to probing with antibody of interest, would it interfere with the antibody binding and the subsequent development of the membrane for...

09 October 2018 506 5 View

Cells adhering at the edges of the culture flask/dishes?

Hi I have noticed one thing in cell culture is that the cells love to attach themselves at the side of the the culture vessels more than at any other part of the flask. Also, the these cells...

07 August 2018 6,159 5 View

HBL-100 Normal or Normal like or transformed?

Hi I am confused regarding the status of HBL 100. Some mention it as 'normal' cell line while others state that it is 'normal like' and another is it is 'transformed'. A clear and precise...

07 August 2018 2,679 3 View

RIPA lysis buffer?

Hi everyone I made a fresh RIPA buffer using foll. composition 1) Tris Buffer 50mM (7.5 pH) 2)150mM NaCl 3)1%NP-40 4)0.5% Sod. deoxycholate 5)Protease inhibitor After lysate was run on SDS-PAGE...

05 June 2018 319 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