CAG promoter vs CMV promoter which one is better for high expression of protein?

Abhijit Chakraborty @Abhijit_Chakraborty8

08 August 2018 1 782 Report

I am constructing one plasmid vector for mammalian expression. I am going to use it for Purification of Protein for structural analysis. Which Promotor is the best one and can give me high yield?

Hua Xiao

The CAG promoter could be a better choice, since it is a synthetic/reconstructed promoter that includes the CMV enhancer/regulatory elements.

Badges
Science method

More Abhijit Chakraborty's questions See All

Which is the best antibody to use in immunoprecipitation (IP) as well as Western Blotting for the study of BRCA1 and BRCA2?

I need to do IP as well as Western Blotting for the endogenous and overexpression BRCA1 and BRCA2 protein. Which is the best one to use?

31 December 2016 6,370 3 View

How can I confirm that a sequence variation is a mutation or Polymorphism?

I found a new variant (a substitution) in exon 20 of BRCA1 gene. This is not reported in HGVS, BIC or any other mutation database. The variant was found after sequencing with both the forward and...

03 April 2016 4,231 14 View

Is there any user friendly protein modelling software rather than MODELLER?

For using homology or comparative modeling of protein three-dimensional structures

31 December 2015 5,234 0 View

Modeller 9.12 shows the following result. How can this be rectified? (File attached)

During protein modelling, Modeller 9.12 shows some problem (see attached file). How can I minimize the error?

05 June 2013 5,035 2 View

How do I calculate the annealing temperature of my PCR reaction?

Tm value of one of my primer is 52.8 (50mM NaCl) and another one is 54.5 (50mM NaCl) as given by the primer making company. Primer Sequence is – i> 5' TGT GTA TCA TAG ATT GAT GCT TTT GA 3' and...

05 June 2013 7,539 49 View

How can I re-concentrate my protein after gel-filtration Chromatography?

I isolated my protein complex from mammalian cells. Purify it using affinity purification methods. but after running chromatography the concentration became very low to go further structural...

01 January 1970 5,267 14 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

Why does my protein refolded to beta sheet during thermal denaturation analysis?

Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...

06 August 2024 1,989 3 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

Low-yield gel extraction problem?

I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...

05 August 2024 9,798 3 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How to Add Missing Water Molecules in Protein-Membrane Simulations?

I have protein-membrane simulations (PDB, PSF, DCD) and have noticed that water molecules near the protein are not visible in the simulations. How can I fix this issue? Is there a way to place the...

04 August 2024 1,200 2 View