Journal:

Article Title: Dislocation of a Type I Membrane Protein Requires Interactions between Membrane-spanning Segments within the Lipid Bilayer

doi: 10.1091/mbc.E03-03-0192

Figure Lengend Snippet: (A and B) Mutation of Gln192 does not affect subcellular localization or MHCI heavy chain binding. (A) HA-US11 cells (top panels) or HA-US11 Q192L cells (bottom panels) were stained with the AF8 mAb (Hochstenbach et al., 1992 ) recognizing the ER chaperone calnexin (left panels, green), biotinylated anti-HA (middle panels, red), and DAPI (blue, all panels). A merge of the left and middle panels (right panels) shows colocalization of calnexin and US11. (B) HA-US11 cells (lanes 1–3) or HA-US11 Q192L cells (lanes 4–6) were labeled for 10 min and chased for 0, 15, or 30 min. A first immunoprecipitation was performed from lysates (1% [wt/vol] digitonin in 25 mM Tris-HCE, pH 7.4, 150 mM NaCl, 5 mM MgCl2) with 12CA5-coupled Sepharose (anti-HA). Bound material was eluted from the beads and denatured by boiling in 1% [wt/vol] SDS with 5 mM DTT. After dilution with NP-40 lysis mix, a second immunoprecipitation was carried out with αHC (top panel) and αUS11 (bottom panel). (C) MHCI-β2m complexes are stable in cells expressing US11 Q192L, but are retained in the ER. Control U373 cells (lanes 1–6) and US11 Q192L cells (lanes 7–12) were pulse-labeled for 15 min and were chased for 0, 1, or 2 h. Immunoprecipitations were performed with antitransferrin receptor (van de Rijn et al., 1983 ; top panels) or W6/32 (Parham et al., 1979 ; bottom panels). Immune complexes were treated with EndoH where indicated. The positions of the EndoH-resistant (EndoHR) and EndoH-sensitive (EndoHS) MHCI heavy chains, US11 Q192L, and β2m are indicated.

Article Snippet: 12CA5 (anti-HA) was coupled to CNBr-activated Sepharose 4 Fast Flow (Amersham-Pharmacia Biotech, Piscataway, NJ) according to the manufacturer's specifications.

Techniques: Mutagenesis, Binding Assay, Staining, Labeling, Immunoprecipitation, Lysis, Expressing, Control

Journal: Cell reports

Article Title: Gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-A K124ub

doi: 10.1016/j.celrep.2021.109924

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-HA (clone 12CA5) , Recombinant Antibody Platform (TAb-IP), Institut Curie , Cat# A-M-M#07.

Techniques: Recombinant, Virus, Plasmid Preparation, Software

Journal: The Journal of Cell Biology

Article Title: Analysis of the Saccharomyces Spindle Pole by Matrix-assisted Laser Desorption/Ionization (MALDI) Mass Spectrometry

doi:

Figure Lengend Snippet: Immunofluorescent staining of HA-tagged spindle pole components with mouse anti-HA mAb 12CA5 ( top ) and rabbit anti-tubulin ( bottom ). Insets, F and I, unfixed diploid GFP-labeled cells at half magnification. Bars, 2.5 μm, except C and G where bar is 2 μm.

Article Snippet: For immunoEM the cells were fixed in formaldehyde for 20 min, the cell wall removed, and then the cells were incubated with either rabbit anti-GFP (a generous gift of K. Sawin, Imperial Cancer Research Foundation, London, UK) or mouse anti-HA mAb 12CA5 and then Nanogold Fab anti-rabbit IgG or anti-mouse IgG (all from Nanoprobes, Stony Brook, NY).

Techniques: Staining, Labeling