Why am I getting 2 bands of 70 KD and 55 KD for the chains of my antibodies on a SDS-PAGE gel?

Dear All, I have purified 3 variants of the monoclonal antibody from CHO cell line expressing them, using 30 KD amicon ultra-4 centrifugal filter devices following by protein G purification,...

06 July 2019 7,989 13 View

Is it practical to repeat infection process to reach better transduction efficiency?

Dear all, I'm working with lentiviral vectors and aiming to overexpress my gene of interest in CHO-k1 cell lines. I have tried several protocols to reach the high titre of the virus, however the...

04 May 2019 852 4 View

Plasmid transfection in HEK293T cell by Calsium Phosphate method ?

Dear All , we are working on gene transfection by Calcium Phosphate method but after 24h when I want to change the cell culture medium of my test and contro cells , I see that tests are not as...

11 December 2018 3,646 8 View

Which CD20 expressing cell line is better?

Hi there , we are working on antibodies against CD20 antigen ( tositumomab, ofatumumab, Obinutuzumab ) and need a cell line which overexpressing CD20 . as far as I am concerned, Daudi , Ramos...

10 November 2018 7,058 5 View

What is the best protocol for lentivirus transfection in HEK293Tcells by "CaHpo4" ?

Hello everyone , I'm trying to do transfection by lentiviral vectors and my packaging cell line is HEK293T. We are going to do calcium phosphate method.I would like to find the best protocol for...

10 November 2018 3,973 4 View

PEI(polyethylenimine) versus CaHpo4 and lipophectamine transfection for virus production in HEK293T cells ?

Hi, I was just wondering if anyone has any personal experience or opinions on using PEI for the production of lentivirus in 293T cells compared to calcium phosphate precipitation and...

09 October 2018 2,286 10 View

Can I use SLA1 instead of XHO1?

Hi there. I want to know is it possible to use SLA1 Enz instead of XHO1? My gene of interest is CD20 And I'm working on PSpax, PMD2G plasmid and PCDH. Any suggestion or comment would be...

08 September 2018 8,697 3 View

Any suggestion or comment in regard of PCR troubleshooting ?

07 August 2018 6,772 11 View

PCR troubleshooting.what should I do to solve the problem ?

07 August 2018 4,152 7 View

What is the problem when we can not see anything on agarose jel electrophoresis ?

I'm trying to clone my specific gene , My sample is 900bp cDNA,after pcr i run it in 1% agarose jel, 100v But I couldn't see any band of marker as well as sample. Any comment and suggestion in...

07 August 2018 5,098 4 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

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02 March 2021 3,060 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

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

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28 February 2021 9,936 2 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View