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

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

How much of cDNA to use for qPCR?

02 March 2016 375 5 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

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 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

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

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 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