How do I prepare pancreas samples for s6k1 and 4ebp1 to avoid over representation of other proteins?

More Kodwo Obeng's questions See All

Would anyone be able to provide me with the book with the title Globalization: A Very Short Introduction?

In view of the research question I posted recently, this book will be of great importance, however, I can't get the soft copy so I was hoping if anyone happens to have it and would really...

24 December 2020 7,199 3 View

How has Globalization affected development in the Global South?

I am a student working on a long essay with my topic as "Globalization; an additional problem or a solution to the problems of the global South countries". In this paper, I seek to discuss the...

26 October 2020 3,966 4 View

Is lentiviral production of antibodies in 293T viable?

Hi, I recently started using lentiviral systems to produce proteins in mammalian cells. Knowing the performance of this impressive system in producing high titers of proteins, I am wondering if...

05 August 2020 5,144 2 View

How do you quantify transwell invasion assays after staining with crystal violet? Anything else apart from counting cells manually?

I am trying to compare the invasiveness among to cell types I am working with via a transwell invasion assay. I am used to counting manually under the microscope, something which I think can be...

19 November 2019 271 5 View

Any suggestions on finding homologous of ncRNA between rats and human?

01 June 2018 1,853 3 View

Please is there an advantage of using agar well diffusion over agar disc diffusion in accessing antimicrobial effects of plant products?

Want to know how efficient well diffusion is over disc diffusion

11 January 2017 9,700 7 View

How can I determine the molecular weight of Lignin using dynamic light scattering?

The lignin is soluble in tetrahydrofuran (THF).

07 August 2014 6,317 3 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

Does any one know how i can prepare liver tissue with transmission electron microscope with good fixation?

i have problem in preparation of liver tissue with bad cell organelles and how i can adjust osmolarity

03 March 2021 8,890 1 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Looking for genomic DNA extraction kits from mouse tissue (centrifuge-based, not spin-column)?

Hello. I am looking to extract genomic DNA from mouse brain and lungs and I need a kit that is not spin-column based because I need to scale up the solution amounts for 400mg tissue (most cap at...

25 February 2021 155 2 View

Has anyone tried to scan fluorescence IHC slides with microarray scanner?

I have an old slide microarray scanner (VIDAR Revolution 4200) and would like to give it a "second life", as IHC scanner array (fluorescence dyes), FISH over tissue or similar. Has anyone tried...

24 February 2021 9,210 1 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 View

What is the best way to manage Western blots?

Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...

23 February 2021 8,609 3 View

Cloudy/high background western blot - can anyone help me?

Hi everyone! I try to detect VDR protein (~48 kDa) using Western Blot. After SDS-PAGE using 10% MP TGX Stain-Free Gel from BioRad I transferred the protein onto an metOH activated PVDF membrane...

23 February 2021 6,266 3 View

Sang Ho Lee

Hi Kodwo.

The kidney is a tough tissues consisting of many tubular structures for the dissociation into single cell suspension. I would suggest the use of mortar and pestle after snap freezing small chopped tissue pieces of the kidney by plunging into LN2 gas for 5 min. Take them into chilled mortar and pestle (at -70C for 1 hour). Wear your thick gloves, and grind them completely to obtain tissue powder. You may need additional LN2 to be poured to make powder without making aggregated clumps.

Take a small amount of the powder (known weight >5~10mg) into a known vol of lysis buffer. Do the remaining routine for your protein contents and make SDS-sample-buffered kidney proteins.

Good luck.

Kodwo Obeng

Thank you very much. I will give up feedback when I am done with your procedure.

Thank you once again.