Issues with my microglia primary culture?

Hi everyone, I'm isolating microglia from P1-2 mice and seeding microglia cells from 3 pups brains/T-25 flask. After seeding, I let this astrocytes-microglia mix culture grow for approx 2-3 weeks,...

06 October 2020 3,916 3 View

Do you have any suggestion for a easy and cheap experiment to quantify Cytokines?

I need to test the stimulation production of TNF alpha in the RAW cell line by LPS. I am looking for an easy and cheap experiment to quantify the serial dilution stimulation in this macrophage...

27 September 2020 5,366 6 View

Good antibodies for D1 and D2 striatal medium spiny neurons?

Hello! I am looking for good antibodies that help in clearly distinguish between D1- and D2-type medium spiny neurons (MSNs) in the striatum, by immunohistochemistry and immunofluorescent...

31 May 2020 5,009 3 View

How to image Methylene Blue stained Nylon Membrane?

I'm performing a Dot Blot and using Nylon Membrane stained with Methylene Blue for a loading control. Does anyone have any advice on how to image this membrane? I've tried using our Li-Cor Odyssey...

22 January 2020 2,178 2 View

Good antibodies for rat M1 and M2 microglia?

Hello! Do you know some good antibodies, suitable for IFL, for differently staining the M2 and M2 microglia phenotypes in rat tissue? I am searching for antibodies specific for RAT because all the...

27 October 2019 7,260 3 View

Anyone can help me about grayanotoxin analytical standards for LC-MS/MS?

I need to buy analytical standards for grayanotoxin I, II and III. I only find the type I. Roberta Merlanti

15 May 2019 2,628 4 View

Can ammonium sulphate acidify sterile media?

I recently did an experiment where I created sterile media by adding 1mM of ammonium sulphate to Murashige and Skoog (MS) Modified Basal Salt Mixture (No Nitrogen) M531. The media was not buffered...

30 October 2017 3,574 4 View

Could someone help me to choose the right concentration and time of neutralizing IL1beta antibody after LPS treatment to block its signalling?

Thanks!

06 October 2016 4,648 3 View

Hi all, Does anyone have experience coding custom non-linear regressions in Sigmaplot?

20 January 2015 4,874 1 View

Is THPS toxic towards methanogenic bacteria? In case it is, is there a "selective (more or less!) biocide against sulphate reducing bacteria?

Tetrakis Hydroxymethyl Phosphonium Sulphate and methanogenic bacteria

21 September 2014 8,375 2 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 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

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 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