Knockdown efficiency reflected in PCR but not western blot?

Hi, I am doing siRNA of a particular gene of interest. however, after several attempts, western blot is not showing evidence of this knockdown, I have tried different antibodies and also longer...

02 March 2019 4,500 4 View

Is it possible to do a Phalloidin staining for Actin on RFP transfected cells?

I am trying to stain for actin filament in RFP transfected cells which already show up at red. Is this possible and any ideas?

04 May 2016 5,643 2 View

What is the best way to seed cells on acrylamide coated coverslips?

i am trying to seed cells on coverslip coated with gels (acrylamide based hydrogels) My main obstacle is my cells aren't settling down as expected so they can grow instead they keep floating, or...

10 November 2015 968 4 View

Any advice on Ethidium Bromide and Calcenin staining for cells seeded on coverslip?

I am trying to stain cells seeded on hydrogel coverslip with EtBr and Calcenin in 2D culture. Any suggestion on the best protocol to follow would be greatly appreciated.

10 November 2015 5,452 3 View

Why does my citrate buffer turn milky during immunohistochemistry?

For my IHC protocol, I am staining for beta-catenin in ovarian sections during antigen retrieval. My samples are heated in citrate buffer for 45 mins. I have also read that signs to show that your...

11 December 2014 6,213 4 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