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Ergic 53

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ERGIC-53 is a membrane-bound protein that functions as a cargo receptor in the early secretory pathway of eukaryotic cells. It is involved in the transport of proteins from the endoplasmic reticulum to the Golgi apparatus. ERGIC-53 acts as a lectin and binds to glycoproteins, facilitating their movement within the secretory system.

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Lab products found in correlation

Sec31a, by BD (3 mentions) Tango1, by Merck Group (3 mentions) Tgn46, by Bio-Rad (3 mentions) Hsp47 and calreticulin, by Enzo Life Sciences (2 mentions) Prolong, by Thermo Fisher Scientific (2 mentions) Vectashield, by Vector Laboratories (2 mentions) Collagen 7, by Abcam (1 mentions) C myc, by Merck Group (1 mentions) Sec23, by Abcam (1 mentions) β tubulin, by Merck Group (1 mentions) Hsp60, by Merck Group (1 mentions) Flag tag (flag), by Merck Group (1 mentions) β actin, by Merck Group (1 mentions) Calreticulin, by Enzo Life Sciences (1 mentions) Hsp47, by Enzo Life Sciences (1 mentions) Tyrosinase, by Santa Cruz Biotechnology (1 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions) Tcs sl confocal microscope, by Leica camera (1 mentions) 8 well chamber slide, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 donkey anti mouse, by Thermo Fisher Scientific (1 mentions)

6 protocols using ergic 53

1

Immunocytochemistry Antibody Protocol

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The following antibodies were used in immunocytochemistry: collagen VII (rabbit anti–human [Abcam]; mouse anti–human [Sigma-Aldrich]), ERGIC-53 (mouse anti–human; Santa Cruz Biotechnology, Inc., and Enzo Life Sciences), Sec31A (mouse anti–human; BD), TANGO1 (rabbit anti–human; Sigma-Aldrich), HSP47 and calreticulin (goat anti–human; Enzo Life Sciences), HA (mouse; BioLegend), SAR1 (mouse anti–human; Abcam), TGN46 (Bio-Rad Laboratories), and mannosidase II (rabbit anti–human; Bio-Rad Laboratories and Merck). Mounting media used in confocal and STED microscopy were either Vectashield (Vector Laboratories) or ProLong (Thermo Fisher Scientific).
Raote I., Ortega Bellido M., Pirozzi M., Zhang C., Melville D., Parashuraman S., Zimmermann T, & Malhotra V. (2017). TANGO1 assembles into rings around COPII coats at ER exit sites. The Journal of Cell Biology, 216(4), 901-909.
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2

Antibody Characterization in Cell Biology

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The following antibodies were used collagen VII (rabbit anti–human [Abcam]; mouse anti–human [Sigma-Aldrich]), ERGIC-53 (mouse anti–human; Santa Cruz Biotechnology, Inc., and Enzo Life Sciences), Sec31A (mouse anti–human; BD), TANGO1 (rabbit anti–human; Sigma-Aldrich; rabbit anti-human in-house), HSP47 and calreticulin (goat anti–human; Enzo Life Sciences), HA (mouse; BioLegend), SAR1 (mouse anti–human; Abcam), β-tubulin (mouse anti-human; SIGMA-Aldrich), β-actin (mouse anti-human; SIGMA-Aldrich), NBAS (rabbit anti-human SIGMA-Aldrich), RINT1 (rabbit anti-human; SIGMA-Aldrich and goat anti-human (Santa Cruz Biotechnology), ZW10 (rabbit anti-human; Abcam), Sec23 (rabbit anti-human/mouse/rat; Abcam), cTAGE5 (rabbit anti-human Atlas antibodies, mouse anti-human Santa Cruz Biotechnology), TGN46 (sheep polyclonal, Bio-Rad), HA (mouse monoclonal, BioLegend; rat monoclonal BioLegend), FLAG (mouse monoclonal, rabbit, SIGMA-Aldrich; goat, Novus) HSP60 (mouse anti-human SIGMA-Aldrich), c-myc (mouse monoclonal, rabbit, SIGMA-Aldrich). Mounting media used in confocal and STED microscopy were either Vectashield (Vector Laboratories) or ProLong (Thermo Fisher Scientific, Waltham, Massachusetts).
Raote I., Ortega-Bellido M., Santos A.J., Foresti O., Zhang C., Garcia-Parajo M.F., Campelo F, & Malhotra V. (2018). TANGO1 builds a machine for collagen export by recruiting and spatially organizing COPII, tethers and membranes. eLife, 7, e32723.
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3

Antibody Panel for Cell Biology Analysis

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Antibodies used in western blotting and immunocytochemistry; Col VII and ATP5A1 (Abcam, Cambridge UK), Syntaxin 18 (Santa Cruz Biotechnology, Dallas, USA), ERGIC-53 (Santa Cruz Biotechnology, Dallas, USA; Enzo Life Sciences, Farmingdale, New York, USA), Sec31A (BD Biosciences), TANGO1, YKT6, c-Myc (9E10), c-Myc and alpha-tubulin (SIGMA-Aldrich, St. Louis, MO, USA), Mannosidase II (Merck, Kenilworth, NJ, USA), TGN46 (abd serotec, Kidlington, UK), HSP47 and Calreticulin (Enzo Life Sciences, Farmingdale, New York).
Santos A.J., Raote I., Scarpa M., Brouwers N, & Malhotra V. (2015). TANGO1 recruits ERGIC membranes to the endoplasmic reticulum for procollagen export. eLife, 4, e10982.
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4

Immunofluorescence Staining of Dendritic Cells

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DC cultured in 8-well chamber slides (Nunc) were fixed (4 % paraformaldehyde), permeabilized (0.1 % Triton X-100), and stained with rhodamine phalloidin (Life Technologies) and DRAQ5 (eBioscience), to label F-actin and nuclei, respectively. The following primary antibodies were used for staining cells: AFP (AbD Serotec), CD206 (BioLegend), CD36, Tyrosinase, EEA-1, LAMP-1, KDEL, ERGIC-53 (all Santa Cruz Biotechnology), golgin-97 (Life Technologies), TGN46 (Sigma), mouse and rabbit IgG isotype controls (R&D Systems), and anti-Alexa Fluor 488 (to amplify the fluorescent signal of Alexa Fluor 488-labeled protein; Life Technologies). The following secondary antibodies were used: goat anti-mouse Alexa Fluor 488 and 555, goat anti-rabbit Alexa Fluor 488 and 555 (all from Cell Signaling Technology), donkey anti-goat Cy3 (Jackson ImmunoResearch), donkey anti-mouse Alexa Fluor-488, and donkey anti-rabbit Alexa Fluor-488 (both Life Technologies). Images were acquired using a Leica TCS-SL confocal microscope.
Pardee A.D., Yano H., Weinstein A.M., Ponce A.A., Ethridge A.D., Normolle D.P., Vujanovic L., Mizejewski G.J., Watkins S.C, & Butterfield L.H. (2015). Route of antigen delivery impacts the immunostimulatory activity of dendritic cell-based vaccines for hepatocellular carcinoma. Journal for Immunotherapy of Cancer, 3, 32.
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5

3D Imaging of Transfected HEK Cells

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Transfected HEK293T cells were plated onto rat tail collagen (EMS, 72295) coated, 22- by 22-mm no. 1.5 coverslips. Cells were fixed in 4% paraformaldehyde, permeabilized with methanol on ice for 10 min, and blocked in 3% bovine serum albumin. Cells were then immunostained with the indicated antibodies: Strep (Qiagen, 1023944), ERGIC-53 (Santa Cruz Biotechnology, sc-365158), Nuclear Pore Complex (Biolegend, 682204), Hoescht 33342 (Invitrogen, H3570), Mouse IgG-AlexaFluor549 (Invitrogen, A11005), and Rabbit IgG-AlexaFluor488 (Invitrogen, A11008). Coverslips were mounted onto glass slides using ProLong Gold Antifade Mountant (Invitrogen, P36934) and analyzed by confocal microscopy (Olympus FV3000RS) using a Olympus UPLAN S-APO 60× OIL OBJ,NA1.35,WD0.15MM objective. The resulting Z-stack was reconstructed and rendered in 3D using Imaris software (Oxford Instruments).
Chen I.P., Longbotham J.E., McMahon S., Suryawanshi R.K., Khalid M.M., Taha T.Y., Tabata T., Hayashi J.M., Soveg F.W., Carlson-Stevermer J., Gupta M., Zhang M.Y., Lam V.L., Li Y., Yu Z., Titus E.W., Diallo A., Oki J., Holden K., Krogan N., Fujimori D.G, & Ott M. (2022). Viral E protein neutralizes BET protein-mediated post-entry antagonism of SARS-CoV-2. Cell Reports, 40(3), 111088.
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6

Signaling Pathway Inhibition Assay

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Pazopanib (GW786034B) and Sorafenib (BAY 43-9006) were obtained from LC Laboratories. The primary antibody ERGIC53 was obtained form Santa Cruz and others primary antibodies pSTAT3 (TYR705, D3A7), STAT3 (124H6), beta-Tubulin, GAPDH were purchased from Cell Signalling while secondary antibodies were purchased from Dianova. Carboxyfluoreszein-Succinimidyl Ester (CFSE) was purchased from Invitrogen, while Hoechst 33342 was provided by Sigma. D-Luciferin Sodium Salt was purchased from PJK GmbH.
Craveiro R.B., Ehrhardt M., Holst M.I., Pietsch T, & Dilloo D. (2014). In comparative analysis of multi-kinase inhibitors for targeted medulloblastoma therapy pazopanib exhibits promising in vitro and in vivo efficacy. Oncotarget, 5(16), 7149-7161.
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