Someone knows how to preconcentrate bacteria density in water samples?

I have low bacteria density in water samples. I thought to preconcentrate them by filtrating a known volume of water (e.g. 500 ml) in 0.2 um Nylon filters, and then suspended bacteria that has...

14 January 2021 772 2 View

Why in standard curve qPCR the most concentrate cDNA sample doesn't work and serial dilutions are correct?

Hi! I did a standard curve of my gene of interest. For this, I started with 100ng of cDNA and did serial dilutions (1:10) of this sample 3 steps. The Ct of this samples are: 100ng --> 32,62...

05 November 2019 3,126 3 View

Problems with transfecting fibroblasts?

Hello I am writing because I am having serious problems transfecting foreskin human fibroblasts with several plasmids at a time. At first we started using the transfection reagent Trans-IT-2020...

01 April 2019 1,276 4 View

Hi, Does anybody know about how to collect fractions of proteins using FPLC AKTA SYSTEM ?

I mean, to different times of elution and specific pikes of the chromatogram. Thank you very much!

30 November 2016 430 2 View

Does anybody know why in the last step of MSP1D1 protein purification this protein precipitates?

05 October 2014 8,744 5 View

How can I stain for oenocytes in L3 larvae (Drosophila)?

I'm trying to establish a baseline of how fed and starved oenocytes look like in L3 larvae on my wild type strains. But I'm having a hard time finding them. This is how I'm doing the experiment:...

10 April 2014 1,987 1 View

Type of immune response using SRBC or NP-CGG (or similar)

Dear all, I have a naive question. When we want to study Type 1 response, we infect mice with influenza for instance, if we want to study Type 2 response we use for instance T. Bruecei. Does...

01 January 1970 220 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

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