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Pcmv ha

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The PCMV-HA is a lab equipment product manufactured by BD. It serves as a general-purpose tool for various laboratory applications. The core function of the PCMV-HA is to provide a controlled environment for specific processes or experiments.

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Lab products found in correlation

C12 ie dap, by InvivoGen (2 mentions) β galactosidase and luciferase activity, by Promega (2 mentions) Hek293 cell, by American Type Culture Collection (2 mentions) Hek293 cells, by BD (2 mentions) Pcmv ha, by Addgene (1 mentions) Pires2 egfp, by BD (1 mentions) Pcmv myc, by BD (1 mentions) Pact vector, by Promega (1 mentions) Pbind vector, by Promega (1 mentions)

4 protocols using pcmv ha

1

Plasmid Construction and Cloning

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HA-WWOX, His-WWOX, His-WW1-WW2, His-WW2-SDR, Myc-WWOX, and Myc-WWOXY33R have already been described [31 (link), 32 (link)]. QuickChange kit was used to obtain HA-WWOXY33R by site-directed mutagenesis (Stratagene, La Jolla, CA, USA). HA-MERIT40 and Flag-MERIT40 were constructed by inserting the appropriate cDNA, obtained by PCR using Myc-Flag-MERIT40 plasmid as template, into pCMV-HA (BD Bioscience Clontech, Palo Alto, CA, K6003-1) and p3xFLAG-CMV-7.1 vectors (Sigma, E4026) respectively. HA-ABRAXAS was constructed by inserting the appropriate cDNA, obtained by PCR using pOZ-N-FH Abraxas plasmid (Addgene, Cambridge, MA, USA, 27495) as template, into pCMV-HA vector. Myc-Flag-MERIT40 (RC202644), Myc-Flag-BRCC36 (RC224289), and Myc-Flag-RAP80 (RC202823) were purchased from OriGene (OriGene Technologies, Rockville, Md, USA). Flag-BRCA1 (52504) was purchased from Addgene, and Flag-BRE (HG16514-NF) from Clinisciences (Montrouge, France).
Taouis K., Vacher S., Guirouilh-Barbat J., Camonis J., Formstecher E., Popova T., Hamy A.S., Petitalot A., Lidereau R., Caputo S.M., Zinn-Justin S., Bièche I., Driouch K, & Lallemand F. (2023). WWOX binds MERIT40 and modulates its function in homologous recombination, implications in breast cancer. Cancer Gene Therapy, 30(8), 1144-1155.
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2

NOD1/NOD2 Signaling Pathway Modulation

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HEK293 cells (ATCC CRL-1573) were obtained from ATCC and were grown in a 48-well tissue culture plates in DMEM containing 10% FBS until ~40% of confluency was reached. HEK293 cells were transfected with a total of 250 ng of plasmid DNA per well, consisting of 25 ng of the reporter construct pNF-κB-luc, 25 ng of the normalization vector pTK-LacZ, and 200 ng of the different combinations of mammalian expression vectors carrying the indicated gene (empty control vector, pCMV-HA-VceC2 , pCMV-HA-TRAF2DN (this study), hNOD1-3xFlag, hNOD2-3xFlag, pCMV-HA-hRip2, hNOD1DN-3xFlag, hNOD2DN-3xFlag or pCMV-HA-Rip2DN3 and pCMV-myc-CDC42DN4 . The dominant-negative form of TRAF2, lacking an amino-terminal RING finger domain5 , was PCR amplified from cDNA prepared from HEK293 cells and cloned into the mammalian expression vector pCMV-HA (BD Biosciences Clontech). Forty-eight hours after transfection, cells were lysed either without any treatment, or stimulated with C12-iE-DAP (1000 ng/mL, InvivoGen) and MDP (10 µg/mL, InvivoGen). After five hours of treatment the cells were lysed and analysed for β-galactosidase and luciferase activity (Promega). FuGene HD (Roche) was used as a transfection reagent according to the manufacturer’s instructions. Cell lines were monitored for Mycoplasma contamination.
Keestra-Gounder A.M., Byndloss M.X., Seyffert N., Young B.M., Chávez-Arroyo A., Tsai A.Y., Cevallos S.A., Winter M.G., Pham O.H., Tiffany C.R., de Jong M.F., Kerrinnes T., Ravindran R., Luciw P.A., McSorley S.J., Bäumler A.J, & Tsolis R.M. (2016). NOD1/NOD2 signaling links ER stress with inflammation. Nature, 532(7599), 394-397.
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3

NOD1/NOD2 Signaling Pathway Modulation

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HEK293 cells (ATCC CRL-1573) were obtained from ATCC and were grown in a 48-well tissue culture plates in DMEM containing 10% FBS until ~40% of confluency was reached. HEK293 cells were transfected with a total of 250 ng of plasmid DNA per well, consisting of 25 ng of the reporter construct pNF-κB-luc, 25 ng of the normalization vector pTK-LacZ, and 200 ng of the different combinations of mammalian expression vectors carrying the indicated gene (empty control vector, pCMV-HA-VceC2 , pCMV-HA-TRAF2DN (this study), hNOD1-3xFlag, hNOD2-3xFlag, pCMV-HA-hRip2, hNOD1DN-3xFlag, hNOD2DN-3xFlag or pCMV-HA-Rip2DN3 and pCMV-myc-CDC42DN4 . The dominant-negative form of TRAF2, lacking an amino-terminal RING finger domain5 , was PCR amplified from cDNA prepared from HEK293 cells and cloned into the mammalian expression vector pCMV-HA (BD Biosciences Clontech). Forty-eight hours after transfection, cells were lysed either without any treatment, or stimulated with C12-iE-DAP (1000 ng/mL, InvivoGen) and MDP (10 µg/mL, InvivoGen). After five hours of treatment the cells were lysed and analysed for β-galactosidase and luciferase activity (Promega). FuGene HD (Roche) was used as a transfection reagent according to the manufacturer’s instructions. Cell lines were monitored for Mycoplasma contamination.
Keestra-Gounder A.M., Byndloss M.X., Seyffert N., Young B.M., Chávez-Arroyo A., Tsai A.Y., Cevallos S.A., Winter M.G., Pham O.H., Tiffany C.R., de Jong M.F., Kerrinnes T., Ravindran R., Luciw P.A., McSorley S.J., Bäumler A.J, & Tsolis R.M. (2016). NOD1/NOD2 signaling links ER stress with inflammation. Nature, 532(7599), 394-397.
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4

Plasmid Construction for Transcription Factor Analysis

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Expression plasmid pcDNA3.1-Mef2c and 3× MEF2-luciferase reporter construct, which contain three upstream tandem repeats of a MEF2 binding site sequence from the desmin gene, were kindly provided by Eric N. Olson.
The Suv39h1 gene was amplified using SF1-SF3 primers (Table 1). Then, the recombinant plasmids were separately digested with Xho I and EcoR I or Sal I and Not I, and ligated into the pIRES2-EGFP (BD Biosciences Clontech, Franklin Lakes, NJ, USA), pCMV-Myc (BD Biosciences Clontech), and pBIND vectors (Promega, Madison, WI, USA), separately.
The Mef2c gene was amplified using MF1 primers (Table 1) designed according to the Sus scrofa Mef2c gene (accession number: NM001044540). Then, the recombinant plasmids were digested with Sal I and Not I, and ligated into the pCMV-HA (BD Biosciences Clontech), and pACT vectors (Promega), separately.
The HP1α gene was amplified using HF1-HF3 primers (Table 1). Then, the recombinant plasmids were digested with Nhe I and Xho I, and EcoR I and Xho I, or Sal I and Not I, and ligated into the pIRES2-EGFP, pCMV-HA, and pBIND vectors, respectively.
Jin W., Shang Y., Peng J, & Jiang S. (2016). Histone H3 Methyltransferase Suv39h1 Prevents Myogenic Terminal Differentiation by Repressing MEF2 Activity in Muscle Cells. International Journal of Molecular Sciences, 17(12), 1908.
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