The domain structure of HEG1 was predicted as follows: signal peptides (exon 1), residues 1-29 proline rich domain (Pro rich) (exon 1), 30-106 serine/threonine rich region (Ser/Thr rich) (exon 2-5), 107-530 Ser/Thr rich (exon 6), 531-630 Ser/Thr rich (exon 7), 631-1086, including poly-serine sequences (polySer), 759772 EGF domain 1 (typical EGF motif) (exon 8), 1087-1125 EGF domain 2 (Ca 2+-binding EGF motif) (exon 9), 1126-1165 unknown region (UN) (this domain may contain a potential proteolytic cleavage site) (exon 10-12), 1166-1273 EGF domain 3 (this domain may form a laminin-type EGF-like domain with a following extracellular juxtamembrane region (EJM) 1) (exon 13), 1274-1318 EJM 1 (EJMs consist of a short loop formed by a disulfidebridge) (exon 14), 1319-1332 EJM 2 (exon 15), 1333-1345 transmembrane domain (TM) (exon 16), 1346-1370 cytoplasmic domain (exon 16-18), 1371-1481. The positions of SNPs in HEG1 of ACC-MESO-4 are indicated with the reference SNP ID number at the bottom.
HEG1 gene sequences were retrieved from GenBank (accession no. (a) Structure of open reading frame of human HEG1 gene. After detection, the membrane was reprobed and treated with anti-β-actin mAb (AC-15). EGFP-transfected HEK293T (HEK293T + EGFP) was used as a negative control. Cell lysates (5 μ L per lane) were resolved by SDS-PAGE under reducing conditions. (k) Representative western blotting (n = 2) of HEG1-transfected HEK293T (HEK293T + HEG1) using mAb SKM9-2. Control siRNA, MISSION siRNA Universal Negative Control (SIC-001) H1097, H2674, and H3671, HEG1 siRNAs copGFP, a control lentiviral particles. Similar results were obtained by 3 independent analyses. Cell lysates (7.5 μ L per lane) were resolved by SDS-PAGE under reducing conditions and detected by western blotting using mAb SKM9-2. (j) Decrease of SKM9-2 antigen by gene silencing of HEG1. The CBB-stained band was then cut from the gel and analyzed by mass spectrometry and Mascot search. The purified sample was resolved by SDS-PAGE. (i) Results of Mascot search for purified SKM9-2 antigen. (h) Western blotting and CBB staining of purified SKM9-2 antigen. Pooled fractions are indicated as a black bar. (d,e,f,g) Purification of SKM9-2 antigen using a Superose 6 Increase 10/300GL, Mono Q 5/50GL, WGA-agarose, or Superose 6 Increase 10/300GL column, respectively. A partially purified sample was treated with glycosidase or proteinase K and resolved by SDS-PAGE under reducing conditions. (c) Elimination of recognition with mAb SKM9-2 by glycosidase treatment. An irrelevant mAb (2D2, mouse IgG1) was used as a negative control mAb. (b) Flow cytometric analysis of SKM9-2 antigen on ACCMESO-4.
Cell lysates (10 μ L per lane) were resolved by SDS-PAGE under non-reducing conditions.
(a) Representative immunoblotting (n = 2) of SKM9-2 antigen in MPM cell lines. Made of faux leather and available in several colors, this travel-friendly case also has slots for your credit cards, ID and boarding pass, and it’s finished with a secure magnetic clasp.MAb SKM9-2 recognizes a sialylated HEG1 on the cell surfaces of mesothelioma cells. This slim passport holder doubles as a vaccine card protector, thanks to the clear window that makes it easy to show your document. If you purchase an independently reviewed product or service through a link on our website, we may receive an affiliate commission. From passport holders that make it easy to show your documents while traveling, to playful movie-inspired cases, see our top picks for the best vaccine card protectors. With that in mind, we’ve rounded up some of the best vaccine card holders for keeping your records safe and handy. But many point out that those four-inch-by-three-inch cards are an inconvenient size for basic billfolds, and others prefer to tote their CDC document in something more stylish or easily portable.
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