Can I leave blot in block buffer for 48 hours?

Is it ok ti leave the blot in non fat milk for 48 hours in +4 degrees or is it better to leave it tbs-t for that extra 24hrs?

01 February 2020 3,899 1 View

Should the HCV Core protein fused with His-tag (without linker) be extracted in the insoluble or soluble protein fraction?

I have infected insect cells SF9 with recomb. baculovirus expressing Core.His and another with GFP. I have extracted the soluble proteins after 3 days and did western blot using anti-his,...

01 February 2020 6,197 4 View

Is it wrong to cut the vector backbone?

I am trying to remove the EGFP insert from pEGFP-N1. I cut the insert using incompatible rest. sites and religate using gap filling PCR and standard ligation reaction. But the transformation on...

04 May 2018 9,335 9 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

How to repeat an operation on an atom selection for multiple times in PyMol?

Hi, I am trying to construct a multi-layer fibril structure from a single layer in PyMol by translating the layer along the fibril axis. For now, I am able to use the Translate command in PyMol...

02 March 2021 4,569 4 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

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28 February 2021 9,936 2 View

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

When is it unnecessary to change organisational culture?

As a former management practitioner, I recall a generally accepted view that changing organisational culture is difficult, so you should only attempt it if the organisational change that you are...

27 February 2021 2,595 8 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

What is the best surface to culture cells isolated from a skin biopsy?

Hello, recently I am having some difficulties with maintaining the cell culture of primary cells isolated from a skin biopsy (normal and malignant). The coating of culture plates with Matrigel (a...

26 February 2021 6,151 2 View