Lipofectamine transfection reagent

Manufactured by Yeasen

Lipofectamine Transfection Reagent is a laboratory product used for introducing genetic material into cells. It is a lipid-based transfection agent that facilitates the uptake of DNA, RNA, or other molecules into cells. The reagent forms complexes with the genetic material, which can then be taken up by the cells, allowing for efficient and reliable gene delivery.

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Lab products found in correlation

Ab252215, by Abcam (1 mentions) Trieasy total rna extraction reagent, by Yeasen (1 mentions) Ab7291, by Abcam (1 mentions) 5 bromouridine, by Merck Group (1 mentions) Mouse s0002, by Affinity Biosciences (1 mentions) Remodelin, by CSNpharm (1 mentions) Complementary dna synthesis supermix, by Yeasen (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) β actin 60008 1 ig, by Proteintech (1 mentions) Sybr green master mix, by Yeasen (1 mentions) Puromycin, by Yeasen (1 mentions)

3 protocols using lipofectamine transfection reagent

Generation of AIMP1 Overexpression and Knockdown Models

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The plasmids containing human AIMP1 cDNA or short hairpin RNA (shRNA) cassettes and mouse AIMP1 cDNA were provided by TranSheepBio (Shanghai, China). Mouse AIMP1 plasmid was transfected to RAW264.7 transiently by Lipofectamine Transfection Reagent (40802ES02, Yeasen, Shanghai, China). The AIMP1‐coding sequence was cloned into the lentiviral vector pTSB with Flag tag (TranSheepBio). AIMP1‐targeted shRNA was cloned into vector pLKO.1 (TranSheepBio). Lentiviruses containing cDNA or shRNA were obtained by co‐transfection of the pTSB‐AIMP1 vector or AIMP1 shRNA vector with packaging vectors (Pspax.2 and PMD2.G, TranSheepBio) into HEK293T cells following the protocol of Lipofectamine Transfection Reagent (40802ES02, Yeasen). MM cells were transfected with lentivirus containing AIMP1 cDNA or shRNA to yield AIMP1 overexpression (AIMP1‐OE) or AIMP1 knockdown (AIMP1‐KD) MM cells. Transfected cells were selected by puromycin (60210ES25, Yeasen). Transduction efficiency was determined by Western blotting (WB).

Wei R., Zhu Y., Zhang Y., Zhao W., Yu X., Wang L., Gu C., Gu X, & Yang Y. (2022). AIMP1 promotes multiple myeloma malignancy through interacting with ANP32A to mediate histone H3 acetylation. Cancer Communications, 42(11), 1185-1206.

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Lentiviral Transduction of SFRS8 in HEK293 Cells

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Plasmids containing human SFRS8 cDNA and SFRS8 shRNA cassettes were provided by TranSheepBio (Shanghai, China). SFRS8 coding sequence was cloned into CD513B1 vector with green fluorescence and Flag tags; SFRS8‐targeting shRNA was inserted into pTRIPZ vector, which was controlled by a DOX‐inducible promoter. Lentivirus packaging and transfection steps were performed as previously described.¹⁶ The expression vector and packaging vector (PLP1, PLP2, VSVG) were co‐transfected into HEK293 cells according to the Lipofectamine Transfection Reagent (YEASEN, Shanghai) method to obtain lentivirus. After 48 h, the virus supernatant was collected, concentrated and stored at −80°C. The transfected cells were screened by puromycin with high transduction efficiency.

Zhang Y., Yu X., Sun R., Min J., Tang X., Lin Z., Xie S., Li X., Lu S., Tian Z., Gu C., Teng L, & Yang Y. (2022). Splicing factor arginine/serine‐rich 8 promotes multiple myeloma malignancy and bone lesion through alternative splicing of CACYBP and exosome‐based cellular communication. Clinical and Translational Medicine, 12(2), e684.

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Antibody-Based Protein Analysis Protocol

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The antibodies were used as following, NAT10 (13365-1-AP, Proteintech; DF12427, Affinity), CDK4 (11026-1-AP, Proteintech), CDK6 (14052-1-AP, Proteintech), CEP170 (18899-1-AP, Proteintech), PARP (9542 S, Cell Signaling Technology), cleaved caspase-3 (9661 S, Cell Signaling Technology), ac4C (ab252215, Abcam), rabbit IgG (7074, Cell Signaling Technology), β-actin (60008-1-Ig, Proteintech), Ki67 (AF0198, Affinity), α-tubulin (ab7291, Abcam), BrdU mAb (MI-11-3, MBL), secondary antibodies included goat anti-rabbit IgG(H+L) HRP (FMS-Rb01, Fcmacs) or mouse (S0002, Affinity). Remodelin was purchased from CSNpharm. Puromycin was from AEPxBIO (B7587), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, M8180) was purchased from Solarbio. Actinomycin D was purchased from MCE (MedChemExpress, HY-17559). 5′-Bromo-uridine (BrU) was purchased from Sigma–Aldrich (850187). Lipofectamine Transfection Reagent (40802ES02), TRIeasy Total RNA Extraction Reagent (19201ES60), complementary DNA synthesis superMix (11123ES10), and SYBR Green Master Mix (11198ES03) were purchased from Yeasen.

Wei R., Cui X., Min J., Lin Z., Zhou Y., Guo M., An X., Liu H., Janz S., Gu C., Wang H, & Yang Y. (2022). NAT10 promotes cell proliferation by acetylating CEP170 mRNA to enhance translation efficiency in multiple myeloma. Acta Pharmaceutica Sinica. B, 12(8), 3313-3325.

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