Why do the mutants aren't expressed in the 293 cells whereas Full-length WT construct is expressed ?

I prepared the recombinant protein X-FLAG tagged and two of its mutant. I have checked the mutant was correctly assembled in the pShuttle vector but cannot discover its expression even after 1...

11 December 2018 3,212 17 View

Overexpression construct: I have 6.5kbp size protein for overexpression tool in 293 cells?

I tried twice with HA-tagging. The construct is made using pShuttle vector but failed in overexpressing the construct.

04 May 2018 6,190 5 View

What is the percentage of mutation rate in a cloning of a recombinant protein? How do you proceed for BL21, once the DH5a positive clone is achived?

Please suggest your opinion!

01 February 2018 3,168 6 View

I want to prepare an overexpression module for my protein with a N-ter HA tag. Can you suggest me the possible strategies?

I have tried by breaking the protein into multiple fragment and using the PCR products to clone one by one. I successfully cloned into the pShuttle vector but I couldnt get the expressed protein,...

01 February 2018 5,966 2 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

Please suggest a good text book for "Research Methodology in Life Sciences". Which book would be good for PhD Scholars?

Please provide Book Title and author name

02 March 2021 9,059 3 View

No title

01 March 2021 3,905 5 View

IHC on mouse liver hepatocytes?

Can anyone help me choose primary antibodies for mouse liver hepatocytes and also in conjunction Cd8 cells. Also Cd3 cells. I want to look at hepatocytes and adjacent immune cells in mice with...

01 March 2021 6,953 1 View

How to avoid ulcers in s.c. injected tumors ?

Recently, we have injected s.c. B16F10 tumor cells in BL/6 mice. At around 12-15th day some animals showed ulcerations in tumors , with a big hole in the middle (it looks like a volcano). We have...

01 March 2021 9,938 3 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

Can random sampling have inclusion criteria?

As random sampling means that everyone in the population have an equal chance to be chosen, can it have inclusion criteria? My research participants have been chosen based on the inclusion...

26 February 2021 6,905 10 View

Looking for genomic DNA extraction kits from mouse tissue (centrifuge-based, not spin-column)?

Hello. I am looking to extract genomic DNA from mouse brain and lungs and I need a kit that is not spin-column based because I need to scale up the solution amounts for 400mg tissue (most cap at...

25 February 2021 155 2 View

What are the pros and cons for mice versus rats?

Hi Everyone, I am designing a pilot study using rodents to assess the effects of simulated space environments (radiation + altered gravity) on microbiome and immune system. I have limited...

24 February 2021 1,825 3 View

Is it harmful to breed for Cre Homozygosity initially?

Hello, I am currently using a Cas9 knock in system that will be expressed in the liver. To do so STOP codon fl/fl Cas9 KI mice were bred with Albumin-Cre mice to generate heterozgous pups, that...

24 February 2021 5,759 3 View