Mef 1 nucleofector kit

Manufactured by Lonza

The MEF 1 Nucleofector Kit is a laboratory equipment product designed for the transfection of primary mouse embryonic fibroblasts (MEFs) using the Nucleofector technology. The kit provides the necessary reagents and solutions to enable efficient delivery of genetic material into MEF cells.

Automatically generated - may contain errors

Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Pcdna3, by Thermo Fisher Scientific (1 mentions) Pcdna3, by Lonza (1 mentions) Amicon ultra centrifugal filter, by Merck Group (1 mentions) Amaxa nucleofector technology, by Lonza (1 mentions) Penicillin, by Nacalai Tesque (1 mentions) Streptomycin, by Nacalai Tesque (1 mentions) X treamgene hp reagent, by Roche (1 mentions) Dulbecco modified eagle medium (dmem), by Nacalai Tesque (1 mentions) 154cf media, by Thermo Fisher Scientific (1 mentions) Lipofectamine ltx with plus reagent, by Thermo Fisher Scientific (1 mentions) Smoothened agonist sag, by Merck Group (1 mentions) Fugene 6 transfection reagent, by Promega (1 mentions) Amaxa nucleofector device, by Lonza (1 mentions) Fetal bovine serum (fbs), by GE Healthcare (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Amaxa nucleofector 2b device, by Lonza (1 mentions)

Close spelling variants of this product

Nucleofector I Nucleofector kit

5 protocols using mef 1 nucleofector kit

Mammalian Expression of Rat MBL-A and Human LH3

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mammalian expression vectors pcDNA3 (Invitrogen) encoding a untagged or a carboxy-terminally FLAG-tagged rat MBL-A [23] (link) and pcDNA3 containing the full length human LH3 or the amino-terminal part of LH3 to the amino acid 290 with a LH3 signal peptide and a c-Myc-tag at the amino-terminus [5] (link),[24] (link) were transfected into LH3^−/− knockout, LH mutant and wild type MEFs using a MEF 1 Nucleofector Kit and Amaxa Nucleofector technology (Lonza). The following day the culture medium was changed to serum free DMEM, penicillin/streptomycin, and 50 µg/ml ascorbic acid, and the cells were cultured for 48 h. The medium was collected and concentrated by Amicon Ultra centrifugal filters (10 kDa MWCO, Millipore).

Risteli M., Ruotsalainen H., Bergmann U., Venkatraman Girija U., Wallis R, & Myllylä R. (2014). Lysyl Hydroxylase 3 Modifies Lysine Residues to Facilitate Oligomerization of Mannan-Binding Lectin. PLoS ONE, 9(11), e113498.

+ Open protocol

+ Expand

Characterization of Wild-type and IRBIT Knockout Mouse Embryonic Fibroblast Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Wild-type (WT) or IRBIT knockout (KO) mouse embryonic fibroblast (MEF) cells have been described previously (Kawaai et al., 2015 (link)). HeLa cells (RRID:CVCL_0030) were obtained from the RIKEN BioResource Center (Ibaraki, Japan) where their identity was checked by Short Tandem Repeat (STR) polymorphism profiling analysis. MEF cells and HeLa cells were cultured at 37°C with 5% CO₂ in Dulbecco’s modified essential medium (DMEM, Nacalai Tesque) supplemented with 10% (vol/vol) FBS, 50 units/mL penicillin, and 0.05 mg/mL streptomycin (Nacalai Tesque). None of the used cell lines were in the list of commonly misidentified cell lines maintained by the International Cell Line Authentication Committee. All cell lines were checked for mycoplasma contamination.
HeLa cells were transfected, according to manufacturer’s instructions, with X-treamGENE HP reagent (Roche Diagnostics) for plasmids and with Lipofectamine 2000 (Thermo Fisher Scientific) for siRNA. For MEF cells, electroporation was performed using MEF 1 Nucleofector Kit (VPD-1004, Lonza) according to manufacturer's instructions.

Bonneau B., Ando H., Kawaai K., Hirose M., Takahashi-Iwanaga H, & Mikoshiba K. (2016). IRBIT controls apoptosis by interacting with the Bcl-2 homolog, Bcl2l10, and by promoting ER-mitochondria contact. eLife, 5, e19896.

+ Open protocol

+ Expand

Hedgehog Signaling Assays in Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

ASZ001 (ASZ, gender: female) BCC cells were cultured in 154CF media (Life Technologies) supplemented with 2% chelated fetal bovine serum, Human Kerytinocyte Growth Supplement (Thermo Fisher), Penn-strep, and 0.05mM CaCl2. Experiments assaying hedgehog signaling carried out in serum-free conditions.
NIH-3T3 (gender: male) and HEK-293T (gender: female) cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS). Experiments assaying hedgehog signaling were carried out in 0–0.2% FBS supplemented with Smoothened Agonist (SAG, Sigma).
Mammalian cell transfection performed using FuGENE® 6 Transfection Reagent (Promega), Lipofectamine® LTX with Plus™ Reagent (Thermo Fisher), and MEF 1 Nucleofector® Kit (Lonza) per manufacturer protocol. Transient transfection mammalian expression vectors are included below. Stable expression produced by piggybac transposition.

Mirza A.N., McKellar S.A., Urman N.M., Brown A.S., Hollmig T., Aasi S.Z, & Oro A.E. (2018). LAP2 Proteins Chaperone GLI1 Movement Between Lamina and Chromatin to Regulate Transcription. Cell, 176(1-2), 198-212.e15.

+ Open protocol

+ Expand

Efficient MEF Transfection with Nucleofector

Check if the same lab product or an alternative is used in the 5 most similar protocols

MEFs were transfected using the Amaxa Nucleofector device (Lonza), program T-020, MEF 1 nucleofector kit (Lonza VPD-1004), and 2 μg DNA, according to manufacturer’s instructions. For all transfections, efficiency was monitored by qPCR.

Goldsmith J., Marsh T., Asthana S., Leidal A.M., Suresh D., Olshen A, & Debnath J. (2020). Ribosome profiling reveals a functional role for autophagy in mRNA translational control. Communications Biology, 3, 388.

+ Open protocol

+ Expand

Generation and Immortalization of Mouse Embryonic Fibroblasts

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary mouse embryonic fibroblasts (MEFs) were generated from 13.5 days old C57BL/6 Neil1^-/-/2^-/- embryos (Gran et al, manuscript in preparation). Limbs were removed from embryos, the tissue was chopped into small pieces and cell suspension was made by pipetting vigorously. MEFs were grown in DMEM medium (Gibco) supplemented with 10% fetal bovine serum (FBS, Sigma), 2 mM glutamine (GlutaMAX, Gibco) and 1x penicillin/streptomycin (Gibco). Cells grown for 4–5 days were frozen at passage 2 in DMEM medium with 20% FBS/10% DMSO. Neil1^-/-Neil2^-/- MEFs displayed the same proliferation rate as wild type MEFs. Experimental procedures were approved by the Norwegian Animal Research Authority. MEFs were immortalized by frequent passaging, using the 3T3 protocol as described previously [20 (link)]. Neil1^-/-Neil2^-/- and Ogg1^-/- MEFs, described previously [12 (link)], were cultured in a humidified incubator at 10% CO₂ in high-glucose DMEM containing GlutaMAX (Invitrogen) supplemented with 10% fetal bovine serum (GE Healthcare). MEFs were electroporated using an Amaxa Nucleofector 2B device and MEF 1 Nucleofector® kit (Lonza, Cat. #VPD-1004) using the T-020 setting as per the manufacturer’s instructions and 3–4 μg of DNA per 1.0 x 10⁶ to 1.5 x 10⁶ cells.

Petrova L., Gran C., Bjoras M, & Doetsch P.W. (2016). Efficient and Reliable Production of Vectors for the Study of the Repair, Mutagenesis, and Phenotypic Consequences of Defined DNA Damage Lesions in Mammalian Cells. PLoS ONE, 11(6), e0158581.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!