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

does any one have protocol for the effecient transfection of Cas9 Protein (RNP) along with gRNA in Hela Or U2OS, MC3T3-L1 cells using lipofectamine 2000, Mirus etc. thank you in advance.

09 October 2019 1,393 2 View

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...

07 August 2016 1,605 13 View

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

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02 March 2021 3,060 3 View

Should we change media after1st 6hrs after plating HUVEC bought from INVITROGEN?it's not recomended by invitrogen.Will DMSO present in stock matter?

We have HUVEC bought from Invitrogen. Cat no -C-003-5c. while first plating the cells we have bought the protocol suggests not to centrifuge the cell with media to remove the DMSO. Should we...

02 March 2021 3,713 2 View

Is there a powerful system for the security of the systems distributed on IoT systems?

Is there a powerful system for the security of the systems distributed on IoT systems?

02 March 2021 3,858 10 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

How Relativistic effects is one reason for lanthanide contraction?

The Lanthanide contraction which is the decrease in ionic radii of the elements in the lanthanide series from atomic number 57, lanthanum, to 71, lutetium, is due to poor shielding of the 4f...

01 March 2021 2,272 4 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 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