How to determine cutoff for an IVD assay?

How to determine LoD, LoQ, Linearilty, Cutoff, Expected values for an IVD assay (infectious disease or physiological state monitoring eg. hormones). Has anyone preformed these, can you explain...

01 February 2020 2,896 3 View

Monoclonal antibodies both capture and detection raised in murine?

Hi, I have come across an IVD quantitative assay for C-peptide where both anti analyte biotin-conjugated antibody which binds to Strep coated wells and HRP conjugated anti-analyte antibody was...

31 December 2019 3,639 3 View

Test for pre-existing antibodies against AAV?

02 March 2019 5,080 7 View

How is enzyme unit defined with respective to substrate concentration?

01 February 2019 358 5 View

After passaging the cells for 24 times, what will be the generation number?

After reviving the cells from -80C and seeding the cells ~1 million in T25 flask when the cells reach 80% confluency then if I passage/subculture the cells for eg. spleen (finite) or CHO or...

06 July 2018 486 7 View

Blue White Screening?

31 December 2017 3,468 7 View

Surfactant vs Defoamer?

Can surfactant and defoamer be used at the same time?. Surfactant (an amphiphilic) works based on reducing the surface tension of water binding thru hydrophobic interaction and hydrophilic...

31 December 2017 3,884 5 View

Final concetration 10% DMSO?

If I had 28million cells/ml (A) and require 11million cells/ml. I add 392.85 ul from (A) + 607.15 ul of 19.4% (since it is 0.6 times more) DMSO. Now final Conc. of DMSO is 10%? Explain with...

11 December 2017 2,952 1 View

Competetive ELISA result interpretation?

Coated antigen conc. used 50ng/100ul antigen1 (is an antibody) conc. 100ng/100ul + purified Mab conc. 1000ng/100ul antigen1 is an recombinant Mab with human Fc region against peptide conjugated...

10 November 2017 4,603 7 View

Why primary ab is diluted in 1% skimmed milk and secondary ab in 1% BSA?

In western blot why do we dilute primary antibody in 1% skimmed milk so that if there are any antibodies against blocking agents they get blocked? but why would they be antibodies against skimmed...

10 November 2017 7,525 7 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

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

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View