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Functional Expression of Human NKCC1 from a Synthetic Cassette-Based cDNA: Introduction of Extracellular Epitope Tags and Removal of Cysteines.

Somasekharan S, Monette MY, Forbush B
PLoS One. 2013 Dec 5;8(12):e82060. doi: 10.1371/journal.pone.0082060. (Link opens in a new window) PubMed

Plasmids from Article

ID Plasmid Purpose
49060pcDNA3.1 Flag Cl-sensitive YFP hNKCC1 WT (NT13)Expresses human NKCC1 with an N-terminal 3xFlag-YFP-(chloride-sensing) tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites. Native hNKCC1 amino acid sequence
49061pcDNA3.1 Flag YFP hNKCC1 N-terminus T,S mutations (NT51)Expresses human NKCC1 mutated Ser and Thr residues in the N-terminus and an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49063pcDNA3.1 Flag YFP hNKCC1 HA-ECL2 (NT931)Expresses human NKCC1 mutant with HA tag inserted into extracellular loop #2 and an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49064pcDNA3.1 Flag YFP hNKCC1 HA-ECL3 (NT933)Expresses human NKCC1 mutant with HA tag inserted into extracellular loop #3 and N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49065pcDNA3.1 Flag YFP hNKCC1 HA-ECL4 (NT935)Expresses human NKCC1 mutant with HA tag inserted into extracellular loop #4 and N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49066pcDNA3.1 Flag YFP hNKCC1 C295S (NT103)Expresses human NKCC1 C295S mutant with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49067pcDNA3.1 Flag YFP hNKCC1 C295A (NT104)Expresses human NKCC1 C295A mutant with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49068pcDNA3.1 Flag YFP hNKCC1 C543M (NT360)Expresses human NKCC1 C543M mutant with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49069pcDNA3.1 Flag YFP hNKCC1 extracellular-cysteineless (NT864)Expresses human NKCC1 mutant lacking extracellular cysteine residues and with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49070pcDNA3.1 Flag YFP hNKCC1 ECL4 deletion (NT865)Expresses human NKCC1 truncation mutant lacking extracellular loop #4 (aa562-582) and with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49071pcDNA3.1 Flag YFP hNKCC1 C630A (NT399)Expresses human NKCC1 C630A mutant with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49072pcDNA3.1 Flag YFP hNKCC1 C630S (NT400)Expresses human NKCC1 C630S mutant with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49073pcDNA3.1 Flag YFP hNKCC1 C723S,C724V (NT501)Expresses human NKCC1 C723S and C724V mutant with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49074pcDNA3.1 Flag YFP hNKCC1 C791S (NT572)Expresses human NKCC1 C791S mutant with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49075pcDNA3.1 Flag YFP hNKCC1 C820S (NT573)Expresses human NKCC1 C820S mutant with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49076pcDNA3.1 Flag YFP hNKCC1 C1052S (NT674)Expresses human NKCC1 C1052S mutant with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49077pcDNA3.1 HA CFP hNKCC1 WT (NT15-H)Expresses human NKCC1 with an N-terminal 3xHA-CFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites. Native hNKCC1 amino acid sequence
49078pcDNA3.1 Flag YFP hNKCC1 TM-cysteineless (NT859)Expresses human NKCC1 mutant lacking cysteine residues within transmembrane domains and with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic,containing convenient restriction sites
49079pcDNA3.1 Flag YFP hNKCC1 cysteineless (NT868)Expresses human NKCC1 mutant lacking cysteine residues and with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49080pcDNA3.1 Flag YFP hNKCC1 intracellular-cysteineless (NT855)Expresses human NKCC1 mutant lacking cysteine residues within intracellular loops and with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites.
49085pcDNA3.1 Flag YFP hNKCC1 (NT17)Expresses human NKCC1 with an N-terminal 3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing convenient restriction sites. Native hNKCC1 amino acid sequence

Antibodies from Article