Brett Seese

6 Questions 1 Answers 0 Followers

Questions related from Brett Seese

Protocol for dilution of cell culture supernatant for sandwich ELISA?

Hi all, I am conducting an ELISA assay to determine the concentration of recombinant protein in the cell culture supernatant after transfection. I have many different transfection conditions, with...

13 October 2021 4,639 3 View

Adding an IgG Fc1 tag to my insert?

I am trying to add an Fc sequence to my insert, and I can't seem to find the sequence anywhere on the internet. I was able to add a FLAG/His tag, but am struggling with this one. Can anyone help...

07 September 2021 6,159 0 View

When to add a N terminal secretion tag?

This is probably a dumb question, but what is the purpose for adding an n terminal secretion peptide to my protein insert during transfection into mammalian cells? Why would I want the protein...

01 September 2021 1,494 3 View

Native signal sequence or addition of new signal sequence?

Hi all, I am transfecting HEK293T cells with SARS-Cov-2 spike protein. I will likely be transfecting the entire sequence after I clone my insert into my plasmid. However, most publications do not...

01 September 2021 4,679 2 View

QPCR to measure transfection efficiency?

I am utilizing a pcDNA3.1 vector and gibson assembly to insert my gene of interest into HEK293T cells using lipid based transfection. I am doing some optimization with a control vector that...

27 August 2021 3,149 3 View

Cloning SARS-COV-2-RBD into pcDNA3.1 vector, how to add tags?

Hi all, I am new to molecular cloning as I had previously only done transient transfection on mammalian cells using RNAi and lipofectamine 3000. So i need to clone the RBD of SARS-COV-2 into my...

24 August 2021 5,861 3 View