Does anyone know how to use image segmentation in image J?

Hi, Im trying to use the image segmentation tool in image J for my image and Iam not getting it correctly to find out the co-localization of two proteins. If anyone has used it, could you please...

09 October 2019 6,877 4 View

Anone facing phototoxicity in IF samples?

Hi, Iam doing IF in HeLa cells and facing a weird problem. I use the standard protocol and when I expose the cell to laser light in the 488 channel, all the fluorescence fades the instant its...

08 September 2018 6,532 4 View

How to design a blocking peptide?

Hi, I want to design a blocking peptide for the C terminus of my protein. If anyone knows how to do it, please help me out. Thanks jaya

04 May 2018 3,735 3 View

Anyone working with connexins?

Hi, Iam encountering a problem in the cell culture. I have hela cells stably expressing connexin 36 attached to GFP on the C terminus. My problem is there is connexin 36 in the internal vesicles,...

11 December 2017 3,842 3 View

Help with cleaving GFP from the protein of interest ?

Hi, Iam working with a protein which is attached to GFP and expressed in Hela cells. I want to do single particle imaging using cryoEM. Im trying to do an IP with GFP antibody and trying to use a...

09 October 2017 2,326 1 View

Protocol for native gel for membrane proteins?

Hi, Does anyone have a protocol for native gel that is working for membrane proteins. Also I have a clarification to be made what is anode and cathode buffer. iam using the tetracell from biorad...

09 October 2017 4,759 2 View

How to represent western blot for paper ?

Hi, I am trying to represent my western blots for paper and have a clarification how to represent the Y axis in the graph. I normalized the test protein with actin and then divided my test protein...

08 September 2017 6,061 5 View

Is anyone working with halotag proteins?

hi, Iam working with a halotag attached to my protein expressed in Hela cells. I was trying to pull down the halotag with magne beads and got nothing in the western. I tried increasing the...

07 August 2017 8,554 3 View

Help with western blot. hello any one there?

hi, I am working with GFP tagged connexin 36 transfected in hela cells. I do a crude membrane preparation and then used chaps to solubliize the membrane for two hrs and run it on a 10% gel. When I...

02 March 2017 9,048 10 View

Should I give the SD in the fold difference and the P value for western blot for publicaiton?

Hi, Im confused on how to present my result for western blot for the paper. I did the experiments thrice by licor and quantified and for each time the machine gave me different values for the same...

09 October 2016 3,329 8 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

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

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02 March 2021 3,060 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

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