Peptide lost during reverse phase HPLC?

Hello; I've dissolved my synthetic peptide in pure acetic acid and tried to purify it with reverse phase HPLC. I got weak peaks during analysis, however I injected at least 5 mg of my peptide!...

02 March 2019 9,491 5 View

How to calculate Kd with flowcytometric analysis?

Hello; I want to calculate Kd value for a FITC-conjugated peptide which specifically binds to a cell receptor. I've studied some protocols, but still confused! Does anyone have any experience or...

11 December 2018 7,119 2 View

Any protocol for solution based ELISA?

Hello; I've isolated peptides against PDL-1 from a phage library, but I can't confirm their binding to target in a solid based ELISA assay. So I want to check whether if I can get a positive...

04 May 2018 1,971 3 View

What is the easiest way to remove SDS following protein purification from gel?

I've purified my recombinant protein from SDS polyacrylamide gel and now I need to remove SDS for further work. Is it possible to do a buffer exchange on amicon filters? could anyone suggest any...

01 February 2017 1,165 4 View

Does anyone know how to justify the sample size in animal studies?

Hi there; I want to investigate anti tumor effects of an antagonist in murine models and I need to know how to determine number of animals in each group such that the results would be...

11 December 2016 1,166 9 View

Is there any way to induce tumorogenesis in murine models using human cell lines?

Hi there; I need to evaluate an anti cancer vaccine against a human protein in murine models.So I should use human cell lines. Is there any convenient way to induce tumorogenesis in murine models...

09 October 2016 1,859 3 View

Is there any convinient assay for in vivo gene expression evaluation?

Hi there; I want to needle inject Balb/C mouse with a plasmid coding for a specific gene and also GFP as a reporter and I need a method for in vivo gene expression assay! Is there any special kit...

06 July 2016 3,577 3 View

Why it's optimal for industrial microorganisms to grow in high temperature?

Hi there; I've read somewhere that growing in high temperatures is an advantage for industrial microorganisms; but unfortunately I couldn't find any reason for this in related articles! Is there...

04 May 2016 3,022 5 View

How can I detect secretary GFP-fused proteins in DMEM supernatant?

I've transfected HEK293-T cells with plasmid expressing for a secretary GFP-fused protein. I would like to know is there any method rather than ELISA test to detect my protein in supernatant using...

04 May 2015 4,653 4 View

What is the optimal concentration of G418 for selection of HEK293-T cells?

I've transfected HEK293-T cells with pCI as a mammalian expression vector containing neomycin gene as a selectable marker. I need to know the most efficient concentration of G418 for cell...

03 April 2015 2,021 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

No title

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

No title

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