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Pbabepuro3 p16flag

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PBabepuro3-p16Flag is a plasmid vector that contains the p16 gene fused to a Flag tag. The plasmid is designed for expression of the p16 protein in mammalian cells.

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Lab products found in correlation

Lipofectamine 3000, by Thermo Fisher Scientific (4 mentions) Pcdna3 myc dnmt1 vector, by Addgene (2 mentions) Lipofectamine 3000, by Addgene (2 mentions) Pspcas9n bb 2a puro px462 v2, by Addgene (1 mentions) Flag p21 wt, by Addgene (1 mentions)

3 protocols using pbabepuro3 p16flag

1

Overexpression of p16, DNMT1 in Breast Cancer Cell Lines

Cited 1 time
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MMTV-PyMT-S2WTP3 cells were infected with lentiviral particles expressing the pHAGE-deltaOA-zsGreen plasmid as previously described6 (link), and infected cells were isolated by FACS.
MDA-MB-453 and BT474 cells were transient transfected with pBabepuro3-p16Flag (Addgene Cat # 24934) using Lipofectamine 3000 (Thermo Fisher) according to the manufacturer’s instructions. 72 hours after transfection cells were selected in puromycin for 48 hours. p16 overexpression was confirmed by anti-FLAG western blots.
For overexpression of DNMT1, MDA-MB-453 cells were transfected with the pcDNA3/Myc-DNMT1 vector (Addgene Cat #36939) using Lipofectamine 3000 according to the manufacturer’s instructions. Cells were treated with abemaciclib 72 hrs after transfection.
Goel S., DeCristo M.J., Watt A.C., BrinJones H., Sceneay J., Li B.B., Khan N., Ubellacker J.M., Xie S., Metzger-Filho O., Hoog J., Ellis M.J., Ma C., Ramm S., Krop I.E., Winer E.P., Roberts T.M., Kim H.J., McAllister S.S, & Zhao J.J. (2017). CDK4/6 inhibition triggers anti-tumor immunity. Nature, 548(7668), 471-475.
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2

Inducible p16 and p21 Overexpression

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For Tet‐inducible overexpression of p16INK4a and p21Cip1 in human primary fibroblasts we used a two‐component transposon system (Figure S1). First, the puromycin resistance gene with a preceding 2A site was integrated into the transposable region of the tet‐controllable transposon plasmid pTOV‐11. This DNA fragment was amplified from the vector pSpCas9n(BB)‐ 2A‐Puro (PX462) V2.0 (Addgene #62987). This was followed by integration of the sequence into the target vector via restriction digestion by BamHI and Crf9I. Integration of the p16INK4a or p21CIP1 cDNA into the target vector followed in another cloning step. The cDNA was amplified from vectors pBabepuro3‐p16Flag (Addgene #24934) and Flag p21 WT (Addgene #16240) and integrated into the transposon vector via NotI and SalI restriction sites.
Brauer E., Lange T., Keller D., Görlitz S., Cho S., Keye J., Gossen M., Petersen A, & Kornak U. (2022). Dissecting the influence of cellular senescence on cell mechanics and extracellular matrix formation in vitro. Aging Cell, 22(3), e13744.
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3

Overexpression of p16, DNMT1 in Breast Cancer Cell Lines

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
MMTV-PyMT-S2WTP3 cells were infected with lentiviral particles expressing the pHAGE-deltaOA-zsGreen plasmid as previously described6 (link), and infected cells were isolated by FACS.
MDA-MB-453 and BT474 cells were transient transfected with pBabepuro3-p16Flag (Addgene Cat # 24934) using Lipofectamine 3000 (Thermo Fisher) according to the manufacturer’s instructions. 72 hours after transfection cells were selected in puromycin for 48 hours. p16 overexpression was confirmed by anti-FLAG western blots.
For overexpression of DNMT1, MDA-MB-453 cells were transfected with the pcDNA3/Myc-DNMT1 vector (Addgene Cat #36939) using Lipofectamine 3000 according to the manufacturer’s instructions. Cells were treated with abemaciclib 72 hrs after transfection.
Goel S., DeCristo M.J., Watt A.C., BrinJones H., Sceneay J., Li B.B., Khan N., Ubellacker J.M., Xie S., Metzger-Filho O., Hoog J., Ellis M.J., Ma C., Ramm S., Krop I.E., Winer E.P., Roberts T.M., Kim H.J., McAllister S.S, & Zhao J.J. (2017). CDK4/6 inhibition triggers anti-tumor immunity. Nature, 548(7668), 471-475.
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