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

Generating Stable Cell Lines with Lentivirus ?

Hello everyone We are working with HEK293T cell line. I would like to know which is the best protocol to Generating Stable Cell Lines with Lentivirus(2nd generation) ? any suggestion or comment...

09 October 2018 3,551 2 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

How to increase a model accuracy ?

dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...

03 March 2021 6,994 5 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Is there any test to determine the cardiovascular risk of patients with obesity? from the perspective of obesity?

We found tests to determine cardiovascular risk taking into account lipid profile, blood pressure, age, history of diabetes, other laboratories. But when weight is involved, there is a paradox. I...

02 March 2021 6,036 1 View

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02 March 2021 3,060 3 View

What SPSS analysis should I run with one IV (with two levels) and two DVs?

Hi, I am trying to figure out which test would be most appropriate to run for the following question: Is there a difference in task blood pressure between smokers and non-smokers? There are two...

01 March 2021 8,235 2 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