What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

Errors in DNA sequences?

I am trying to better understand the scope of DNA replication and sequencing errors, e.g. 1. I have seen similar error rates of 10e4 to 10e5 for cell & instrumental DNA replication,...

24 February 2021 4,397 3 View

What can I do if DNA OD260/280 is either 1.6 or higher than 3?

26 January 2021 7,830 4 View

How do you multiple impute item scores and perform a linear regression on factor scores?

I am developing a questionnaire and first performing an exploratory factor analysis. After I have the final factor structure, I plan on regressing the factor scores on some demographic covariates....

18 January 2021 883 8 View

How do I design an extra long linker?

I'm trying to insert a linker between two proteins that is almost 100 amino acids in length, does anyone have any experience designing such a linker? I plan to order a gene block from IDT and...

17 December 2020 7,812 1 View

Is the fluorescent protein must be totally photobleached in FRAP assay?

Hi I have tried to measure the mobility of a protein of interest by the FRAP assay. The protein was fused with GFP. I overexpressed the fusion protein in Hela cells. However, when the area of the...

14 December 2020 6,567 1 View

Why am I unable to coat oleate-capped NaCeF4:Gd,Tb nanoparticles with mesoporous silica shell?

My oleate-capped NaCeF4:Gd,Tb NPs have been synthesized via the thermal decomposition method. 1mmol of NPs has been dispersed in 20 ml of cyclohexane. I have tried dissolving 1 ml of NP in...

01 December 2020 4,713 2 View

Primer specificity check with primer blast individually or together?

Hi, When checking my designed primers for specificity, what is the difference if I run BLASTn for each primer individually versus together (ie. forward primer separated by 20 "N" then reverse...

03 November 2020 1,421 4 View

How to perform statistic analysis for age range between two cohorts?

Hi, I was reading the paper (MicroRNA Expression, Survival, and Response to Interferon in... ). In Table 1, I'm wondering how did the authors calculate a p-value of 0.29 for Age? The age in Table...

20 October 2020 2,276 5 View

Intron contetion and primer selection?

Dear all I am trying to clone a gene, which only contains only "one intron" in the middle part. I treat the extracted RNA with DNAse and use verso cDNA kit, so the DNA contamination ie...

10 September 2020 7,022 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

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