How much should we add primary antibody into 20 mL of 1% Skim milk for Western Blot?

Hello, Everyone! I currently start doing western blot experiment, but I am not really sure how much should I add primary antibody into 20mL of 1% Skim milk. Here is what I am doing: * I make 1%...

10 November 2019 5,213 9 View

Can anyone explain why do we get diffused bands in western blot? and any trouble shooting techniques?

Hello, Everyone. I have some questions related to western blot as I am new to western blot. I tried to detect the LC3B, however I got diffused bands as I have attached in the file (Please find the...

10 November 2019 5,796 13 View

Any recommended handling method for neuroblastoma SH-SY5Y cells?

Good afternoon, I have some question related to the handling method for neuroblastoma cells SH-SY5Y. First, I grew the cells with EMEM medium (10% FBS and 1% Penicillin/Streptomycin). The cells...

07 August 2019 8,334 9 View

Any good handling methods for SH-SY5Y cells?

Hello, Everyone! I have some questions related to the SH-SY5Y cell handling. I grow my SH-SY5Y cells in cell culture dish with EMEM (FBS+, P/S+)cell culture media. One day after cell plating,...

06 July 2019 1,306 1 View

Could anyone recommend me the standard protocol of the design of (microRNA or gene) primers for RT-PCR as a reference?

I am looking for a standard protocol of how to design a good primer for RT-PCR. Could anyone recommend me the standard protocol of the design of (microRNA or gene) primers for RT-PCR as a reference?

05 June 2019 6,715 6 View

Why the expression pattern in Real-time PCR keeps changing for the same primer?

Good morning, Everyone! I have some questions related to Real-time PCR. I have checked the expression of a microRNA (microRNA primer) in monolayer and spheroid cells. The first time I run the...

05 June 2019 4,282 13 View

Which RNA extraction method provides better result between the extraction with QIAzol and that with TRIzol?

Firstly, I have extracted RNA samples by using QIAzol, and then I also tried to use TRIzol to extract RNA samples. Can anyone tell me which RNA extraction method is better between the extraction...

10 November 2018 9,089 5 View

Possible Target genes or Primers for miRNA?

Hi everyone, Could you tell me how to find possible target genes or primers for a specific microRNA when we perform RT-PCR? My experiment is I transfected cells with a specific miRNA, then I...

08 September 2018 4,462 7 View

What somehow makes WST-1 cell proliferation experiment not working ?

Hi Everybody, I have some questions related to WST-1 cell proliferation assay. I did a WST-1 assay experiment with my pancreas cancer cell line in 96-well plate. First, I plated the cells in...

08 September 2018 7,614 0 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

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02 March 2021 3,060 3 View

Ester cleavage conditions?

Hi all, I need to cleave the ester in Ethyl 2-amino-1H-imidazole-5-carboxylate to make the carboxylic acid and retrieve the product from solution to use in downstream applications of linking to...

01 March 2021 8,766 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