Why should skim-milk be used for blocking buffer in western blots?

I have just performed my first western blot and none of my targeted proteins were detected. The only thing visible on the membrane was the prestained ladder. Someone suggested that my blocking...

21 August 2018 5,787 4 View

How to grid downscale CMIP5 climate data grid to 0.5 degree using Matlab ?

08 July 2018 2,235 2 View

Merging many to 1 in SPSS version 25?

I have 2 datasets one is the adult in the houshold and the other is the child(ren). I'm trying to merge the child data to the adult data however there may be more than 1 child to an adult (many...

10 May 2018 4,946 6 View

Why would LB-low salt medium be prefereable to normal LB medium?

I've come across a protocol that says to use low salt LB broth (10g/L tryptone, 5g/L NaCl, 5g/L yeast extract) for bacterial growth curves. Why would it be preferable to use a low salt medium...

18 February 2018 2,048 2 View

Why is the deletion of bcsQ using lambda red recombination unsuccessful?

I have been unsuccessful in knocking out bcsQ in Pectobacterium carotovorum ssp. brasiliense using a three-step PCR method. Plasmids used: pKD4 and pKD46 bcsQ is MinD homologue part of the...

17 July 2016 1,533 2 View

Has anyone seen this strange FSC-A versus FSC-H pattern?

I recently ran an experiment where I divided a mouse tumor into several wells of a 96-well plate. Each well was stained with a different panel of antibodies. In only one of the wells, I got the...

22 February 2016 4,653 6 View

Does anyone have suggestions about cell density, DIV and surface preparation for hippocampal neuron culturing for micro electrode array?

27 November 2014 4,362 2 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

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

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02 March 2021 3,060 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

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 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

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 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