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The ER-Golgi intermediate compartment (ERGIC) marker ERGIC-53 is a mannose-specific membrane lectin operating as a cargo receptor for the transport of glycoproteins from the ER to the ERGIC. ERGIC-53 can also be used for studying aspects of protein trafficking in the secretory pathway.
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Western blot analysis of extracts (35µg) from 9 different cell lines by using ERGIC-53 monoclonal antibody (OTI1A8).
Immunohistochemistry staining of paraffin-embedded carcinoma of human prostate tissue using ERGIC-53 monoclonal antibody (OTI1A8).
Immunohistochemistry staining of paraffin-embedded human prostate tissue within the normal limits using ERGIC-53 monoclonal antibody (OTI1A8).
ERGIC-53 monoclonal antibody (OTI1A8) immunofluorescent staining of COS7 cells transiently transfected by pCMV6-ENTRY ERGIC-53.
Flow cytometric analysis of Jurkat cells, using ERGIC-53 monoclonal antibody (OTI1A8) (red), compared to a nonspecific negative control antibody (blue).
Immunofluorescence microscopy of HEK293A cells shows Surf4 accumulates in and around ERESs.(A) Fluorescent signal for endogenous Surf4 (green) was strongest at punctate structures positive for ERES marker Sec23 (red). Note additional Surf4 fluorescence in weblike structures surrounding ERES. (B) HA-Surf4 signal (green) was observed within the ERGIC (ERGIC-53, red). (C) HA-Surf4 (green) showed only low levels of colocalization with cis-Golgi marker, giantin (red). (D) Mutation of proposed COPI recycling motif by replacement of two of three near-carboxy-terminal lysines to alanines (HA-Surf4-AAK, green) increased colocalization with cis-Golgi marker, giantin (red). (E) Newly synthesized HA-Surf4 (green) was found at low levels in the rER (Sec61 marker, red). (F) Surf4 (green) did not colocalize with chaperone, calnexin (red), in the quality control domain. HEK293A cells were transfected with wild-type HA-tagged Surf4 plasmid (B, C, E), or Surf4KO HEK293A cells were transfected with carboxyl-terminal di-lysine mutation, HA-Surf4-AAK (D), 18 hr prior to fixation. Bars = 5μm. The cells in each panel are shown 3 times, first with organelle marker, then Surf4 and final panel (with magnified insert) showing overlap. Alexa Fluor secondary antibodies were used for detection. Images were obtained using an LSM 780 (Carl Zeiss) confocal microscope (488 and 561 nm excitation lines; 500–560 and 600–660 nm capture) and Zeiss Axio Imager Z1 with Apotome 2 (single Z stack slice). Images were analyzed using Zeiss Zen software. AAK, alanine-alanine-lysine; COP, coat protein complex I; ERES, endoplasmic reticulum exit site; ERGIC, endoplasmic reticulum–Golgi intermediate compartment; HA, hemagglutinin; HEK293A, human embryonic kidney cell line 293A; rER, rough ER; Surf4, surfeit locus protein 4.
Image collected and cropped by CiteAb under a CC0-1.0 license from the following publication: Surf4 (Erv29p) binds amino-terminal tripeptide motifs of soluble cargo proteins with different affinities, enabling prioritization of their exit from the endoplasmic reticulum. PLoS Biol (2018)
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