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
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Questions related to Transfection
Hi, I'm having some trouble running a basic immunoprecipitation experiment. I'm trying to pull-down actin from RAW cells, but all I get is a smear throughout the entire lane with no visible or...
05 April 2022 2,123 9 View
Hello everyone, I am an R beginner and I would like to do a Fisher's exact test 2x2 for each gene on my list (see attached file) to statistically check if there are any differences in...
04 April 2022 7,000 23 View
I did ApoTox-Glo Triplex Assay to asses viability/cytotoxicity and apoptosis on three prostate cancer cell lines (LNCaP, 22Rv1 an PC-3) stimulated with high concentration of docetaxel and...
03 April 2022 1,093 3 View
Hello, I am Mahmuda, now I am working with DG44 cell culture. So far, my cell culture viability improved to 90%. However, before transfection, my cell viability decreased to 70%. I am very...
30 March 2022 480 1 View
I would like to learn my students how to perform a co-immunoprecipitation experiment, coupled to western blot of primary antigen and the co-precipitant. Does anyone have a good suggestion for...
28 March 2022 7,134 3 View
Hello everyone, I wanted to ask if anyone has been able to successfully transfect HEK293 cells with the BioRad Gene Pulser XCell and the pre-set settings used for it. I've often ended up with...
28 March 2022 8,133 1 View
I used shRNA plasmid for the knockdown of gene. Firstly, package the virus, then transfect the target cells and then puromycin selection to get the stable cells. After selection of stable cells i...
28 March 2022 7,694 1 View
Hello, I am trying to measure caspase activity in CHO cell samples using the Flurochrome-labeled inhibitor of caspases assay (FLICA). I want to look at ethanol (or another checmical) induced...
25 March 2022 5,809 2 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 5,972 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 2,204 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 1,736 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 5,027 1 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 7,567 1 View
I guess Quinine works the other way around, blocks Cx36 but not so much of Cx45.
17 March 2022 9,089 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 5,269 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 7,478 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 6,111 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 1,753 1 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 9,155 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 1,149 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 9,792 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 6,534 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 5,332 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 2,561 0 View