Does anyone have a protocol for coating microscope slides with lipids?

05 May 2017 1 4K Report

Hi all --

We are hoping to coat microscope slides in the lipid DOPC. Does anyone have protocol for this?

Thanks for the reference Volodymyr. Do you know if the stream of nitrogen gas is required if the lipid is ordered as a powder (instead of under chloroform)?

Badges
Science method

More Samantha Jeschonek's questions See All

Determining RNA quality after RNA-IP for RIP-Seq. Bioanalyzer ?

Does anyone have experience assessing RNA quality after RNA-immunoprecipitation? My downstream goal is RNA-seq (RIP-seq), and I want to check the quality of the RNA before proceeding to library...

01 February 2018 981 3 View

Tips for protein resolution: 65 vs 75 kD?

Due to limited sample, I'm trying to maximize each SDS gel/Western Blot that I run. In an ideal world, I'd like to resolve a 40 kD, 65 kD, and 75 kD protein on the same gel. I prefer to cut my...

10 November 2017 6,290 4 View

Does anyone know of a good resource for mass spec analysis workflow?

Hello all -- Does anyone have a good paper/book/website that goes through the analysis of (label-free) mass spec data starting from .raw files? I'm specifically looking for resources that use...

07 August 2017 6,768 9 View

My protein was detectable by Western Blot but not mass spec...ideas?

We did an IP experiment and sent the results for mass spec. I confirmed by western blot (on the same sample) that some positive controls were present -- including the antigen. The mass spec...

04 May 2017 1,637 5 View

Remove glycerol from antibodies?

Can anyone recommend a method or product to remove glycerol from small volumes of affinity purified antibody? We are using Dynabead M270 Epoxy beads to couple antibodies to beads. This kit...

03 April 2017 7,162 12 View

Is sonication of lysate required for crosslink-RNA-IP?

I'm doing a crosslink RNA IP. Briefly, my cells are crosslinked with formaldehyde prior to cell lysis and immunoprecipitation. After eluting my sample and reversing the crosslink, RNA is extracted...

02 March 2017 7,467 3 View

RT-qPCR: Very different amplification curves between samples. Short linear phase? Drooping plateau?

I'm having some trouble with my RT-qPCR. Some of my amplification curves (delta RN vs cycle #) look as expected, but others have a distorted curve. High starting background that drops, followed...

09 October 2016 8,573 7 View

Does anyone have experience with RNA-IP: enrichment over input qPCR validation?

Hello - I have a question regarding the best way to setup a (two step) RT-qPCR experiment. I have performed an RNA-IP and using qPCR want to determine if a specific RNA is enriched over (1) input...

07 August 2014 1,454 9 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

Do I need specific antibodies to stain for a fusion protein?

I am staining some brain sections stored in cryoprotectant that express a Histone H2B- GFP fusion protein that were generated ~10 years ago. I know I need to enhance signal with an anti-GFP...

07 August 2024 5,338 2 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

How to determine positive-stained cells in FACS? Use isotype or unstained control?

To compare positive and negative cell populations in flow cytometry, should I compare unstained cells with antibody stained cells? Or with the isotype control? Most papers show comparison with...

06 August 2024 6,728 6 View

Can I multiplex a mouse monoclonal and rat primary for IHC (mouse brain tissue)?

Hi all, I was just wondering if anyone has experience with multiplexing a mouse monoclonal primary and a rat primary. I'm trying to multiplex by incubating them in the same well but was told by a...

06 August 2024 9,710 1 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How to Add Missing Water Molecules in Protein-Membrane Simulations?

I have protein-membrane simulations (PDB, PSF, DCD) and have noticed that water molecules near the protein are not visible in the simulations. How can I fix this issue? Is there a way to place the...

04 August 2024 1,200 2 View