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

What does the last number in cell line name mean?

Hi there; Does anyone one know what's the difference for example between SK-MEL-28 and SK-MEL-37? what does that number mean?

11 December 2016 3,576 0 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

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

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

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 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

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 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