pCMV6-AN-His Mammalian Expression Vector

Snippet: build 11, Bio-Rad) was used to prepare the images. 4.6. BIDC1 cDNA Construct’s Western Blotting A BIDC1 cDNA construct containing the coding sequence of transcript 1 (NM_001714) was purchased from ORIGENE (RG218807). The BICD1 cDNA was subsequently cloned into the open reading frame of the pCMV6-AN-His mammalian expression vector (ORIGENE, #PS100011) using SgfI and MluI. Next, site-directed mut

Snippet: Liposome Tethering Assays. Human E-Syt1SMP-C2AB (residues 93–634) was amplified by PCR and cloned using AscI and NotI sites into the pCMV6-An-His vector (Origene) for Expi293 cell expression. Human TMEM24SMP-C2 (36-414) and C2CD2SMP-C2 (39-405) were cloned using NheI and XhoI sites into pET28a vector (Novagen)

Snippet: Coding sequences of mouse INPP5K was cloned into a modified pCMV6-AN-His vector (OriGene), which has a GFP tag inserted just after His tag. Active site mutation D210A was constructed by site directed mutagenesis

Snippet: Plasmids expressing murine caspase-1 and caspase-3 with C-terminal Myc and DDK (FLAG) tags (pCMV6 entry) and empty vectors for expression of N-terminal Myc-DDK (pCMV6-AN-Myc-DDK) and N- and C-terminal His-tagged proteins (pCMV6-AN-His and pCMV6-AC-His, respectively) were obtained from OriGene Technologies. …

Snippet: The regions coding for residues 93–1,104 (E‐Syt1cyto), 93–941 (SMP‐C2ABCD), 93–634 (SMP‐C2AB), 315–1,104 (C2ABCDE), 624–973 (C2CD), and 936–1,104 (C2E) of human E‐Syt1 were amplified by PCR and cloned using AscI and NotI sites into the pCMV6‐An‐His vector (OriGene) for Expi293 cell expression