Is there any difference between using EDTA and Trypsin to detach cells for Flow Cytometry?

I want to use FAC to study integrins on the cell surface and I've heard some people claim that Trypsin can induce integrin degradation. I'm just wondering if anyone has seen any significant...

06 July 2019 4,878 3 View

How to know if a protein actually exists in the cells?

Im working on a protein. We have our cell line and we suspect that the protein is present there. However, we don't have any commercial antibody for this protein so we cannot do any western blot to...

09 October 2018 3,707 4 View

How do you quantify the number or the signal intensity of intracellular vesicles?

It sounds like a quite straightforward question, however I find it's actually something very difficult to quantify. The problem is that cells have different thickness, and so when you take the...

03 April 2018 189 0 View

What analysis/software/algorithm can I use to analyse if cells in a group are moving randomly or are undergoing some sort of patterned movement?

At the moment, I'm trying to analyse a difficult set of data that I've been thinking but really don't know how to do it. So basically, i have two populations of cells and I notice one of them...

01 February 2018 3,581 2 View

Can't detect the increase of the overexpressing protein using retroviral vector?

So I have made several stable melanoma cell lines expressing a protein of interest using retroviral vector. The cells survive antibiotic selection while those without the construct are all dead....

11 December 2017 571 4 View

Multiple comparison for nested-ANOVA analysis?

08 September 2017 1,132 1 View

Side effect of using GM6001 (a Matrix metalloprotease inhibitor) in cell culture?

04 May 2017 184 3 View

Statistically comparing samples of different conditions to the control?

For example, if you have 4 samples A, B, C, D: A = Control sample B, C, D are samples at different conditions. Assume all assumptions are met. Can you just simply use t-test Can you just simply...

03 April 2017 3,843 8 View

Transforming biological data with n = 3?

03 April 2017 1,748 29 View

Setting threshold when analysing image data?

I'm analysing some imaging data but having a bit of a problem. When the images were taken, some of them have uneven shading. I want to set a threshold to analyse the area certain features within...

01 February 2017 914 6 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

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

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

Markers of mitochondrial activity in FFPE tissues?

Hello everybody, Does someone have any experience with assessing/extimating mitochondrial activity on FFPE tissues? Precisely, no TOM20, ATP5B, etc: we are trying to get an approximative idea of...

24 February 2021 4,618 4 View

Can a Golgi-stained dendritic spines picture processed by Image J be directly used for publication?

24 February 2021 3,348 2 View

Is there an appropriate CD8 antibody for IHC-P for staining human breast cancer tissues?

Hello, We are planning on staining Formalin-fixed paraffin breast cancer tissue sections (FFPE) with CD8 Ab via immunofluorescence (Tissue IF). Based on our existing protocol and imaging...

24 February 2021 9,578 4 View

Reagents for histology slides?

Dear all, I am planning to study histology and want to do that at home. I have only stained a blood smear with Giemsa but now want to try a wider range of animal tissues. What reagents will I...

24 February 2021 5,430 3 View

Are there lab markers/pens resistant to Roti-Histol?

Hi, I'm looking for a pen/marker to write on the smooth, transparent (non-frosted) part of microscopy slides which does not get washed away by alcohols and most importantly Roti-Histol. I want to...

24 February 2021 9,902 5 View