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

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

Is there any dedicated site or a research article where i can find the status of p53 (WT or mutated) in various cell lines?

I need to know the p53 status of HSC-T6 and HK-2 cell lines. Can anyone help?

03 April 2018 469 3 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

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

01 March 2021 8,087 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

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 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

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View