The introduction of DNA into a recipient eukaryote cell and its subsequent integration into the recipient cells chromosomal DNA. | Contact experts in Transfection to get answers
1,621 views 3,440 posts
Questions related to Transfection
I need to know what the Autofluorescence Eliminator Reagent for IHC is diluted in and in which concentration. It comes in 50ml hence I do not think it can be used pure. Thank you for your assistance!
25 March 2022 5,028 0 View
Hi all, It's my first time culturing NK cells, not only mine but also the first time it is being done in my lab so I am kind of on my own. Does anyone have a step-by-step protocol they could...
24 March 2022 5,980 1 View
We have recently generated CAR-Jurkat and CAR-pbmc cells， however， although the initial CAR% is around 80%， after 2 days of culture in vitro， the CAR expression dramatically falls to 20% or...
24 March 2022 2,212 2 View
I am trying to stain organoids and where I am getting results with different staining protocols, I do noticed difference between protocols, especially when it comes to permeabilization component....
23 March 2022 1,745 3 View
I am doing a fairly large panel of transfected genes of interest and also comparing Latrunculin-A actin depolymerization to cytochalasin-D actin depolymerization. I am hoping to skip some...
22 March 2022 5,037 1 View
Dear community! I am about to measure cytosolic Calcium of Vero cells treated with Staurosporin or ionomycin as ionophore. In all of my experiments my Control (untreated cells) was already really...
21 March 2022 1,021 0 View
I've been piloting clonogenic survival assays to look at long-term effects of of drugs in colon cancer cell lines (SW620, SW480, HT-29). My initial pilot assays in 12-well plates were successful...
18 March 2022 7,583 1 View
I guess Quinine works the other way around, blocks Cx36 but not so much of Cx45.
17 March 2022 9,101 2 View
Hi all, I have a question about my lip3000 transfect neurons. I transfected my primary neurons with a control(mRFP) plasmid. Then I did immunostaining with primary proteinX antibody and Alexa 488...
16 March 2022 5,281 0 View
Hi all, The target of my experiment is to purify EGFP tagged protein of interest expressed in mammalian cells (transient transfection). I need information on what kind of columns can be used to...
16 March 2022 7,492 2 View
I'm building a custom pooled gRNA library, for a CRISPR-KO screen. Would add a bunch of positive controls, but also negative (since my screening is not huge, I intend to include around 1/3 of...
16 March 2022 5,021 2 View
I experience high cell death rates and I am looking for a buffer that can be used instead of the P3 buffer for the siRNA transfection of murine nTreg using the nuecleofactor Lonza system. Thanks!
14 March 2022 6,119 2 View
We are planning to transfect HEK293T cells with plasmid that carries shRNA sequence against the gene of interest, grow a large batch and let them produce shRNAs, and then isolate shRNAs. But the...
11 March 2022 1,764 1 View
Company medium is a high price, is the a better alternative i.e, DMEM/F12 ?
10 March 2022 4,481 2 View
Hi everyone, I'm trying to visualize dendritic spines in iPS-derived neurons by transfecting a GFP-actin plasmid. My goal is to achieve a low-efficiency transfection to analyze single neurons...
08 March 2022 9,166 0 View
I am planning to use CRISPR/Cas9 to endogenously label my protein of interest with a tag, I then want to use these cells (U2OS cell line) for imaging. Would it be better to use a fluorescent tag...
08 March 2022 1,158 0 View
The plasmid molarity required for my transfection is 2.75µM. I don't know the initial molarity of my plasmid, I know just the lyophilized micrograms (5 µg) I have now in my tube and the kb of my...
07 March 2022 9,805 3 View
Hello, I am recently studying BMDM (bone marrow derived macrophage), want to transfect a gene into BMDM cells, with common liposome transfection can not go in, want to ask whether there are other...
04 March 2022 6,542 3 View
We are using the Neon Transfection system with Epi5 Vectors. The fibroblasts will be at passage 3 when transfected and they are from an adult donor. After transfection they will be placed in...
02 March 2022 5,346 1 View
I would like to nucleofect human BJ1 hTERT immortalised fibroblasts with a RNP complex made of Cas9 and a single crRNA+Tracer RNA, using the Amaxa 4D cell nucleofector device. Is there a standard...
02 March 2022 2,575 0 View
It was left out for overnight in room temperature. Can we use it for experiments or is it expired? SDS says it should be stored in -20. Has anyone had the same issue and tried to use it?
02 March 2022 9,404 1 View
Essentially, we have various HCC cell lines that we plan to do a migration assay using corning transwell 8.0 um pores w/ matrigel. Utilized a chemoattractant of 5% FBS/DMEM serum as the...
01 March 2022 5,748 2 View
Any help would be appreciated
28 February 2022 6,574 6 View
Hello friends, We are trying to depolarize neuronal cell by kcl, research used this way didn't elaborate much about preparing kcl. I read that we can prepare kcl medium, but I don't have a clear...
26 February 2022 3,735 4 View