How to tranfect Cas9 (RNP) protein into Hela cells using lipofectamine, crispmax or Mirus?

More Muhammad Zahoor's questions See All

Any one have experienced with western blotting for Hif1a?

Hi, i am having trouble with immunoblotting for HIF1a, and would like to have some tips from you guys. i have tried 4-12% SDS gel, dried/fast and wet transfer both, using Anti-Hif1a from R&D...

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DAB kit for goat primary ab?

Hi, i am interested to do immunohistochemistry (DAB staining) on mouse tissue, and looking if some one can suggest me any good kit. I do have primary antibodies from goat against the protein i am...

11 December 2015 4,034 4 View

How to seperate or purify the double allele KO cells from single allele KO cells, using CRISPR/Cas9?

I am using CRISPR/Cas9 system to KO a gene and the plasmid carries GFP as label. However, I am not sure how would I be able to differentiate or purify the double allele KO cells from single allele...

05 June 2015 2,321 8 View

Are you familiar with the HUVEC cells tube formation assay?

I am interested to perform HUVEC cells tube formation assay. Currently I am culturing HUVEC cells in Medium200+LSGS and can I use the same medium for tube formation assay? Please suggest a...

03 April 2015 2,170 3 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

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 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

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

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01 March 2021 6,142 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

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Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

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Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

Sandra Lagauzère

Hi Muhammad!

well, I am not sure you will have a detailed protocol for each cell line you ask.

As tech support for the Viromer CRISPR (another reagent made for RNP delivery), I would suggest to first try the different manufactuerers' protocols and to follow their optimization guidelines. Each new CRISPR experiment needs to be calibrated empirically. There is no ONE FITS THEM ALL protocol.

for more details about our polymer-based reagent, you can go here:

https://viromer-transfection.com/crispr

and

Poster Delivery of Cas/gRNA RNP using the VIROMER® platform

Chne Jing

I'm Chne Jing.

I've successfully transfected gRNA plasmid in Hela cells using lipo-2000 and lipo-3000. Just do it as it's described on the protocol, usually contained in the transfection kits.