Is there any methods to improve mouse ULK2 expression and transfection efficiency?

Hi everyone, I am recently overepressing mouse ULK2 in HEK-293T cells. However, the expression level is much lower than other genes constructed with the same vector. So, is there any methods to...

05 June 2018 6,145 2 View

Is there any validated ULK2 antibody?

Hello everyone, I am recently searching an validated ULK2 antibody. However, the molecular weight by the indicated antibodies from the data sheet is around 112 kD. In my opinion, though the 112 kD...

05 June 2018 9,013 4 View

Why the efficiency of Flag antibody IP is very low?

Hi everyone, Recently, I am performing IP using anti-Flag Ab(M2, F3165) for Flag-ULK1 overexpression cells. But the efficiency is very low. Even if the input accounts for 1% of the whole cell...

07 August 2016 1,823 4 View

Which post-translation modification will change or increase the apparent electrophoresis mobility of protein?

I am recently performing Western Blot for a protein. After treatment, the apparent electrophoresis mobility increases detected by its phosphorylation antibody. Moreover, the other protein...

03 April 2016 2,576 4 View

Could a small molecule target another small molecules?

03 April 2016 4,038 5 View

Why do different exposure times get different results for Western blot?

11 December 2015 431 8 View

Should I boil protein samples containing GPCR or Dopamine receptors for Western Blot?

11 December 2015 2,925 11 View

Why RNAi resistant cell lines cannot rescue the phenotype?

hi everyone, I am recently performing RNAi rescue assay to validate the siRNA for gene/protein A specificity. I construct cell lines expressing wild type gene/protein A and RNAi...

10 November 2015 133 10 View

How can I get functional GPCR in cell lysate?

I am recently performing MST assay to detect the relationship between GPCR and chemical compound. I performed the MST assay using the cell lysate in HeLa cells overexpressing the GFP-tagged-GPCR...

08 September 2014 1,491 13 View

Fugene 6 transfection troubleshooting in HEK 293T

Recently I am performing the transfection assay using Fugene 6 reagent in HEK 293T cell. The plasmid includes the EGFP gene. Even I use the vector including only GFP without the fusion gene, the...

08 September 2014 4,183 7 View

How to compare two groups with only two measurements?

Hiiiii everyone! I have an enquiry on statistical analysis. I was looking for many forum and it's still cannot solve my problem. I want to compare means of two groups of data but only with two...

03 March 2021 8,796 3 View

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02 March 2021 3,060 3 View

How does Scientist/RG community evaluates a case study article and the impact of it on a researcher career?

An article that is published as a case study in the SJR listed journal. How much value is added to a researcher career? How it is evaluated by a scientific researcher/University admission...

02 March 2021 9,661 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

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28 February 2021 9,936 2 View