Non animal derived proteins for blocking in immunoassays?

What would you recommend? It should be cheap and of constant / defined composition, comparable to defatted milk powder (NFDM). Thanks!

31 December 2017 4,384 7 View

Reduction resistant dimers in SDS-PAGE?

Dear Colleagues, I'm working on a recombinant protein (isolated from 3T3 mouse fibroblast supernatants) that behaves a little strange in analytics: Purified by IMAC (it has a His6-Tag) and gel...

11 December 2016 7,444 4 View

Unconjugated Fluorescein in Immunofluorescence?

anyone knows by chance, which mammalian cellular structures it is staining? Just curious. Thanks!

05 June 2016 844 0 View

Sephadex G200 vs Sephacryl S200?

Dear experts, I am planning a clean-up step in a protein purification process where I actually thought of using Sephacryl S200. In the stock room, I found some Sephadex G200 and thought, I'd give...

11 December 2015 9,742 6 View

How do I reduce imines in the presence of aldehydes-alternatives to NaBH3CN?

Dear Colleagues, Any suggestions for alternatives to cyanoborohydride (in respect of less toxic, cheaper, suitable for aqueous settings)? It's about the reductive amination of a polysaccharide...

02 March 2015 6,920 10 View

What is the usual nature of impurities in synthetic peptides?

Dear Colleagues, peptide synthesis companies offer different purities at different prices.What actually defines the purity? Several things come into my mind: - sequence errors (i.e. usually...

01 February 2015 4,456 7 View

How do I remove a stuck rotor from a Biofuge pico centrifuge?

Dear Colleagues, I happened to get a pretty soiled used centrifuge of this type and simply can't get the rotor out for a thorough cleaning of the machine. Of course, the screw has been removed and...

31 December 2014 5,798 9 View

Do you have any suggestion for GLP-1 ELISA?

Dear Colleagues, I'm in search of a ELISA to detect Glucagon Like Peptide 1 (GLP-1) in tissue culture supernatants and serum. Requirements: - detects total GLP-1 (active and inactive, N-terminal...

10 November 2014 9,443 2 View

Can anyone help with a reciepe for a stabilizer for HRP conjugates?

Dear colleagues, Has anyone got a recipe for said buffer/solution to share? I have a secondary antibody conjugated to HRP that I'd like to store at higher dilution in the fridge/freezer. Probably...

07 August 2014 5,619 6 View

Are there any implications of short dyades and palindromic sequences?

Dear colleagues, I have designed a construct for eukaryotic expression. Before I submit it for synthesis, I would like to find out about the possible implications of 3 (unintended) features that...

07 August 2014 3,295 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