Michael Moles

7 Questions 4 Answers 0 Followers

Questions related from Michael Moles

Can you freeze EdU-stained tissues ex vivo (e.g. in OCT compound) for subsequent fluorescence IHC staining?

We wish to use the Click-iT EdU Plus Kits to visualise cell proliferation of mouse splenocytes ex vivo. One method of detecting and visualising EdU incorporation is fluorescence IHC. However,...

29 April 2019 6,373 1 View

Combining conjugated (direct) and non-conjugated (indirect) primary antibodies for IHC - which step?

I wish to probe for two targets using a conjugated and non-conjugated primary antibody (raised in different species) for my IHC experiment. Question is, when would I include the conjugated...

30 November 2018 1,877 3 View

Does anyone have experience in imaging murine plasmacytoid dendritic cells (pDCs)?

We wish to perform immunocytochemistry with FACS sorted murine pDCs. Our issue concerns finding a suitable method for attaching cells to the slide. We have tried the cytospin method, but were not...

21 August 2015 789 2 View

Is it possible to combine direct and indirect staining for flow cytometry?

I wish to detect a protein of interest by combing direct and indirect staining for flow cytometry. The idea would be to stain mouse splenocytes and lymph node cells with fluorochrome-conjugated...

06 August 2015 5,754 4 View

How can I determine the GC content of a gene of interest?

I wish to amplify a DNA fragment by PCR for molecular cloning. As such, I would like to find out the GC content of my gene of interest (cDNA template). Can ensembl or NCBI provide me with this...

01 April 2015 7,525 4 View

Does anyone have experience in generating murine Plasmacytoid Dendritic Cells (pDC) from FLT3 Ligand-supplemented bone marrow cultures?

I have been trying to generate murine pDC by supplementing bone marrow cultures with FLT3 Ligand. I seed 1 x 10^6 cells/ml in 6-well plates, using 200ng/ml of FLT3-L. When harvesting the cells...

26 November 2014 3,357 8 View

Are there any tips for homogenizing a single mouse inguinal lymph node for total RNA isolation?

I wish to isolate total RNA (Trizol method) from a single mouse inguinal lymph node weighing ~10mg. After trying thorough physical homogenization with a glass mortar and pestle, I noticed that...

20 August 2014 665 2 View