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

Role of phosphoric acid in colloidal coomassie staining

In homemade colloidal gel stains based on Coomassie Blue G250, 4..12% phosphoric acid are added as last step to promote colloid formation. How does it work: Is it "just" pH or do phosphate...

02 March 2016 9,959 2 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

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

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

No title

02 March 2021 3,060 3 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

How to simultaneously perform multiple biochemical assays?

I am a research scholar working on heavy metal stress on plants. I will be doing biochemical characterisation( protein, carbohydrate, proline, antioxidant enzymes and many more assays) at interval...

01 March 2021 6,999 3 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

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

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View