Jetprime buffer

Manufactured by Polyplus Transfection

Sourced in France, United States

JetPRIME buffer is a buffer solution designed for use in transfection experiments. It is intended to facilitate the delivery of nucleic acids into cells.

Automatically generated - may contain errors

Lab products found in correlation

Jetprime reagent, by Polyplus Transfection (7 mentions) Jetprime transfection reagent, by Polyplus Transfection (6 mentions) Jetprime, by Polyplus Transfection (3 mentions) Control sirna, by Santa Cruz Biotechnology (2 mentions) Pspax2, by Addgene (2 mentions) Jetprime sirna transfection reagent, by Polyplus Transfection (1 mentions) Glutamine, by Merck Group (1 mentions) Hek293t, by Takara Bio (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Interleukin 2 (il 2), by Thermo Fisher Scientific (1 mentions) Phytohemagglutinin (pha), by Merck Group (1 mentions) Hepes, by Merck Group (1 mentions) M csf, by R&D Systems (1 mentions) Pmdlg prre, by Addgene (1 mentions) Pmd2 g, by Addgene (1 mentions) Prsv rev, by Addgene (1 mentions) Interferin, by Polyplus Transfection (1 mentions) Polybrene, by Qiagen (1 mentions) Blasticidin, by Thermo Fisher Scientific (1 mentions) Lenti x concentrator, by Takara Bio (1 mentions)

39 protocols using jetprime buffer

HMGCR siRNA Knockdown Assay

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Cells were plated in 6-well plates for 24 h, then cells were transfected with transfection reagent INTERFERin (#409-10, Polyplus, France), Jet PRIME Buffer (#712-60, Polyplus, France) and HMGCR (ELAVL1, KHSRP, DHX36, HNRNPD and ZFP36) siRNA or siRNA-negative control (Jet PRIME Buffer: 200 μL; JetPRIME: 2 μL; 20 μmol/L siRNA: 2.5 μL for per well) for 24 h. The siRNA sequences used in the study are listed in Supporting Information Table S1.

Zhang W., Pan X., Xu Y., Guo H., Zheng M., Chen X., Wu H., Luan F., He Q., Ding L, & Yang B. (2023). Mevalonate improves anti-PD-1/PD-L1 efficacy by stabilizing CD274 mRNA. Acta Pharmaceutica Sinica. B, 13(6), 2585-2600.

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Transfection and Analysis of NBC Transporters

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Human HEK293T (H.T), lung fibroblast MRC5, lung adenocarcinoma A549 cells were purchased from ATCC (Washington, DC, USA). Cells were cultured in Dulbecco’s Modified Eagle Medium containing 10% fetal bovine serum (FBS) and 100 U/mL penicillin-streptomycin antibiotics at 37 °C in 5% CO₂/95% air. Human NBCe1-B (an electrogenic form of sodium bicarbonate cotransporter, encoded by SLC4A4) and NBCn1 (electroneutral form of sodium bicarbonate cotransporter, encoded by SLC4A7) clones were provided by Shmuel Muallem (National Institutes of Health, Bethesda, MD, USA). Plasmid DNAs (1 μg/μL) were mixed with 200 μL Jet Prime Buffer (B200225, Polyplus-transfection, Illkirch-Graffenstaden, France). Transfection reagent (4 mL) (21Y0910L1, Polyplus-transfection) was incubated with mixed Jet Prime Buffer and DNA for 10 min in the dark. Incubated DNAs were transferred into H.T-cultured plates, and all procedures were performed in accordance with the protocol of the manufacturer (Polyplus-transfection). MRC5 cells were used for determination of cell viability, and A549 cells were used for determination of native NBC activity. S0859 (18497, Cayman, Ann Arbor, MI, USA) was used for the specific inhibition of NBC activity.

Ji M.J., Son K.H, & Hong J.H. (2022). Addition of oh8dG to Cardioplegia Attenuated Myocardial Oxidative Injury through the Inhibition of Sodium Bicarbonate Cotransporter Activity. Antioxidants, 11(9), 1641.

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STAT1 Knockdown in NCI-H1299 Cells

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NCI-H1299 cells were plated at 50% confluency in 6-well plates for 24 h, then cells were transfected with transfection reagent JetPRIME (Polyplus, #114-15), Jet PRIME Buffer (Polyplus, #712-60) and STAT1 (CaMKII) siRNA or siRNA-negative control (Jet PRIME Buffer (Polyplus, #712-60); 200 µL, JetPRIME 2 μL, 20 μM siRNA: 2.5 μL for per well) for 24 h. The siRNA sequences used in the study are provided in Supplementary Table S1.

Pan X., Li R., Guo H., Zhang W., Xu X., Chen X, & Ding L. (2021). Dihydropyridine Calcium Channel Blockers Suppress the Transcription of PD-L1 by Inhibiting the Activation of STAT1. Frontiers in Pharmacology, 11, 539261.

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Transfecting H292 cells with c-Myc siRNA

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H292 cells (1.5 × 10⁵) were seeded in 6-well plates for 24 h. Then, cells were transfected with transfection reagent JetPRIME (Polyplus-transfection, #114–15, Illkirch, France), Jet PRIME Buffer (Polyplus-transfection, #712–60), and c-Myc siRNA or siRNA-negative control (Jet PRIME Buffer (Polyplus-transfection, #712–60) 200 μL, JetPRIME 2 μL, 20 μM siRNA 2.5 μL for per well) for 24 h (Ding et al., 2019 (link)). The siRNA sequences used in the study are provided in Supplementary Table S1.

Chen X., Du Q., Guo H., He Q., Yang B, & Ding L. (2022). Bafetinib Suppresses the Transcription of PD-L1 Through c-Myc in Lung Cancer. Frontiers in Pharmacology, 13, 897747.

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Noxa siRNA Transfection in DLD-1 and HCT116 Cells

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DLD-1 and HCT116^wt cells were seeded in 6-well plates (6×10⁵ cells with 2 ml medium per well) and cultured at 37°C for 24 h. A mixture containing 110 pmol Noxa siRNA (cat. no. sc-37305; Santa Cruz Biotechnology, Inc.) or control siRNA (cat. no. sc-36869; Santa Cruz Biotechnology, Inc.), 200 µl jetPRIME^® buffer (Polyplus-transfection SA) and 4 µl jetPRIME^® siRNA transfection reagent (Polyplus-transfection SA) was vortexed for 10 sec, spun down and incubated for 10 min at room temperature. The mixture was subsequently added to each well of the 6-well plate and incubated at 37°C for 24 h. Cells were then harvested for further experiments.

Li X.L., Zhou J., Xia C.J., Min H., Lu Z.K, & Chen Z.R. (2021). PRIMA-1met induces autophagy in colorectal cancer cells through upregulation of the mTOR/AMPK-ULK1-Vps34 signaling cascade. Oncology Reports, 45(5), 86.

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Differentiation and Maintenance of Human Monocytic and Adherent Cell Lines

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Human pro-monocytic cell lines U937 and U1.1 (NIH reagents program) were maintained in RPMI 1640 (Cell Clone, CC3014) with 10% FCS (Gibco Life Technologies, 10270) at 37°C in 5% CO2, humidified incubator. Adherent cells HEK293T (Clonetech) and TZM-bl (NIH reagents program) cells were maintained in DMEM (Cell Clone, CC3004) and 10% FCS, as described above. Monocytic cells were differentiated into macroPHAges by treating them with phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich Co, P8139) at 10 ng/ml for 24 h. Adherent cells were seeded in tissue culture-treated Petri-plates or six-well plates in complete DMEM media overnight and transfection and titration experiments were carried out as explained below. Reagents like HEPES (N-2-hydroxyethylpiperazine-N9-2-ethane sulfonic acid; Sigma- Aldrich Co, H3784) sodium bicarbonate (Sigma- Aldrich Co, S5761), glutamine (Sigma- Aldrich Co, G3126) etc. were used for media preparations. Transfection reagents Jet Prime and Jet Prime buffer, (Polyplus™, 114-07), Retro-concentrin (Clonetech Laboratories Inc., RV100A-1), MCSF (R&D systems, 216-MC-025) and used at a concentration at 50 ng/ml for activating MDM’s. IL2 (eBiosciences, 14-8029-63) was used at a concentration of 20 units/ml. PHA (Sigma Aldrich Co, L8902-5mg) was used at 1 µg/ml.

Sharma V., Makhdoomi M., Singh L., Kumar P., Khan N., Singh S., Verma H.N., Luthra K., Sarkar S, & Kumar D. (2020). Trehalose limits opportunistic mycobacterial survival during HIV co-infection by reversing HIV-mediated autophagy block. Autophagy, 17(2), 476-495.

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Retroviral and Lentiviral Particle Generation

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The pBabe, pBabe-myc-BirA*-LMP1, and pBabe-LMP1-BirA*-HA
retrovirus particles were generated by Jetprime transfection of HEK293T cells in
10 mm dishes with pBabe vectors and packaging plasmids pMD2.G (Addgene, 12259)
and pSPAX2 (Addgene, 12260). Five μg of each construct DNA were used per
plate with 30 μL of Jetprime and 500 μL of JetPrime buffer
(Polyplus) was mixed and incubated at room temperature for 25 minutes. The
DNA-Liposome complexes were then added dropwise to each place. Media was
collected and replaced with fresh DMEM at 48, 72, and 96 hours
post-transfection, centrifuged at 1,000 × g for 10 minutes, filtered
through a 0.45 μM filter into sterile tubes, and frozen at
−80°C until use. To make shRNA lentiviral stocks, HEK293-T cells
were transfected with expression plasmids (pLenti X1 Syntenin-1 shRNA, pLenti X1
ALIX shRNA, or pLenti X1 scramble shRNA) and the packaging plasmids pMD2.G
(Addgene; #12259), pMDLgpRRE (Addgene; #12251), and pRSVRev
(Addgene; #12253) to produce retroviral particles for transduction.
Media was collected and stored as described for pBabe viruses. All packaging
plasmids were gifts from Didier Trono.

Rider M.A., Cheerathodi M.R., Hurwitz S.N., Nkosi D., Howell L.A., Tremblay D.M., Liu X., Zhu F, & Meckes DG J.r. (2018). The Interactome of EBV LMP1 Evaluated by Proximity-Based BioID Approach. Virology, 516, 55-70.

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Lentiviral Transduction of Cells

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Lentiviral particles were packaged using HEK293FT cells seeded at a density of 250,000 cells per well in 6-well culture plates. In brief, 1 μg of pLKO.1-Puro TRC plasmid containing either non-targeting control shRNA (5’-CAACAAGATGAAGAGCACCAA-3’) or ATF4-targeted shRNA (5’-GCCTAGGTCTCTTAGATGATT-3’) was combined with 1 μg of packaging plasmids psPAX2 and pMD2.G, mixed at a molar ratio of 2:1. The plasmid mix was diluted in jetPRIME buffer (Polyplus Transfection, 89129-924) and transfection reagent, following the manufacturer’s protocol. pMD2.G and psPAX2 were gifts from Didier Trono (Addgene plasmid # 12259, #12260). Lentiviral supernatants were collected at 24 and 48 h post-transfection, supplemented with 8 μg/ml Polybrene, filtered through 0.45-μm nitrocellulose filters, and stored at −80 °C. Cells were transduced with lentivirus and were selected for viral infection via addition of 2 μg/mL of puromycin for 72 hours.

Baniulyte G., Durham S.A., Merchant L.E, & Sammons M.A. (2023). Shared gene targets of the ATF4 and p53 transcriptional networks. bioRxiv.

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MCF-7 Cells Overexpressing GABARAPL1

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MCF-7 control cells (C) and MCF-7 Flag:GABARAPL1:6His (GABARAPL1) cells were obtained previously in our laboratory following transfection with pcDNA3.1 control, pcDNA3.1-Flag-GABARAPL1-(His)6 [47 (link)]. MCF-7-Flag:GABARAPL1-G116A:6his (GABARAPL1 G116A c1 and c2) cells were obtained in the same way following transfection with pcDNA3.1-Flag-GABARAPL1-G116A-(His)6 vectors. The cells were cultured in Dulbecco's minimum essential medium (DMEM) (PAA, E15-891) supplemented with 100 μg/ml penicillin/streptomycin (PAA, P11-010) and 10% fetal bovine serum (FBS) (PAA, A15-101) in a 5 % CO₂ incubator at 37°C. To inhibit autophagosome/lysosome fusion, cells were incubated for 2 h in complete medium supplemented with 100 nM bafilomycin A1, 40 μM chloroquine or 50 mM NH₄Cl. To induce autophagy, cells were incubated in EBSS for 4h at 37°C. To inhibit proteasome degradation, cells were cultured in complete medium supplemented with 2 μM MG132 for 16 h.
For transient transfection, 2 μg of pGFP, pGABARAPL1-GFP, pGABARAPL1-G116A-GFP, 200 μl Jetprime Buffer and 4 μl Jetprime reagent (Polyplus transfection, 114-07) were used per reaction according to the manufacturer's protocol.

Poillet-Perez L., Jacquet M., Hervouet E., Gauthier T., Fraichard A., Borg C., Pallandre J.R., Gonzalez B.J., Ramdani Y., Boyer-Guittaut M., Delage-Mourroux R, & Despouy G. (2017). GABARAPL1 tumor suppressive function is independent of its conjugation to autophagosomes in MCF-7 breast cancer cells. Oncotarget, 8(34), 55998-56020.

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Transient Silencing of ANXA1 in ECC-1 Cells

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For transient ANXA1 silencing, transfection was performed in 6-well plates using the jetPRIME reagent (PolyPlus Transfection) with, alternatively, ON-TARGETplus Human ANXA1 (301) siRNA (J-011161-07-0010; Dharmacon) or control siRNAs (SIC001; Sigma-Aldrich) on shC1GALT1 and SCRAMBLE ECC-1 cells, according to established protocols [41 (link), 42 (link)]. Briefly, 2.5 × 10⁵ cells were transfected with 22 pmol siRNA using 2 µl of JetPRIME Transfection reagent and 100 µl of JetPRIME buffer (PolyPlus Transfection). Then, 48 h after transfection, cells were analyzed by PCR, qPCR and WB. Alternatively, 24 h after transfection cells were used for indicated cell-based assays as above.

Montero-Calle A., López-Janeiro Á., Mendes M.L., Perez-Hernandez D., Echevarría I., Ruz-Caracuel I., Heredia-Soto V., Mendiola M., Hardisson D., Argüeso P., Peláez-García A., Guzman-Aranguez A, & Barderas R. (2023). In-depth quantitative proteomics analysis revealed C1GALT1 depletion in ECC-1 cells mimics an aggressive endometrial cancer phenotype observed in cancer patients with low C1GALT1 expression. Cellular Oncology (Dordrecht), 46(3), 697-715.

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