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Tgn46

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The TGN46 is a lab equipment product offered by Bio-Rad. It is a device used for the detection and analysis of specific proteins in biological samples. The core function of the TGN46 is to perform Western blot analysis, a widely used technique in molecular biology and biochemistry.

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Lab products found in correlation

Sec31a, by BD (5 mentions) Tango1, by Merck Group (5 mentions) Vectashield, by Vector Laboratories (4 mentions) Flag tag (flag), by Merck Group (4 mentions) Ergic 53, by Santa Cruz Biotechnology (3 mentions) Alexa fluor 488 conjugated donkey anti mouse igg, by Thermo Fisher Scientific (3 mentions) Calreticulin, by Enzo Life Sciences (3 mentions) β actin, by Merck Group (3 mentions) Hsp47 and calreticulin, by Enzo Life Sciences (2 mentions) Lamp1, by Cell Signaling Technology (2 mentions) Procollagen 7, by Abcam (2 mentions) Sec16a, by Merck Group (2 mentions) Grasp55, by Proteintech (2 mentions) Hrp conjugated secondary antibody, by Jackson ImmunoResearch (2 mentions) On targetplus sirna, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Dharmafect 1 transfection reagent, by Thermo Fisher Scientific (2 mentions) Tgn46, by Thermo Fisher Scientific (2 mentions) Vti arrayscanner, by Thermo Fisher Scientific (2 mentions) Ssea 3, by STEMCELL (2 mentions)

Spelling variants of this product

Sheep anti-TGN46 (16 citations) Anti-TGN46 (9 citations) Sheep anti–human TGN46 (5 citations) TGN46 antibody (3 citations) Polyclonal sheep anti-TGN46 (2 citations) Anti-Myc (X) and anti-TGN46 (2 citations) Sheep anti-TGN46 (AHP500GT) (2 citations) TGN46 (AHP500) (2 citations)

22 protocols using tgn46

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

Comprehensive Immunofluorescence and Western Blot Protocol

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We used the antibodies in parentheses to the following antigens: ATG9A (Abcam, catalog #108338), FLAG M2 (Sigma, F1804), PAK3 (Abnova, PAB2300), SPTLC2 (Abcam, ab23696), GFP-HRP (MACS, 130091833), actin-HRP (Sigma, A3854), beclin 1 (Cell Signaling, 3738), ATG7 (Cell Signaling, 8558), ATG5 (Cell Signaling, 12994), ATG12 (Cell Signaling, 4180), ATG16 (Cell Signaling, 8089), LC3 (Cell Signaling, 3868), LAMP-1 (Cell Signaling, 9091), SNAP29 (Abcam, 138500), gp41 (NIH AIDS Reagent Program, 2F5), TGN46 (Bio-Rad, AHP500G), anti-HIV immunoglobulin (NIH AIDS Reagent Program, HIV-Ig), Nef (NIH AIDS Reagent Program, 2949), α-tubulin (Sigma, T5168), VSV-G (Sigma, V5507), Alexa Fluor 488-conjugated donkey anti-rabbit IgG (Invitrogen, A21206), Alexa Fluor 488- conjugated donkey anti-mouse IgG (Invitrogen, A21202), Alexa Fluor 555-conjugated donkey antirabbit IgG (Invitrogen, A31572), Alexa Fluor 555-conjugated donkey anti-mouse IgG (Invitrogen, A31570), Alexa Fluor 555-conjugated donkey anti-sheep IgG (Invitrogen, A21436), HRP-conjugated donkey anti-rabbit IgG (GE Healthcare, NA934V), and HRP-conjugated sheep anti-mouse IgG (GE Healthcare, NXA931).
Mailler E., Waheed A.A., Park S.Y., Gershlick D.C., Freed E.O, & Bonifacino J.S. (2019). The autophagy protein ATG9A promotes HIV-1 infectivity. Retrovirology, 16, 18.
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3

Antibody Characterization Protocol

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The following antibodies were used procollagen VII (rabbit anti–human [Abcam]; mouse anti–human [Sigma-Aldrich]), Sec31A (mouse anti–human; BD), TANGO1 (rabbit anti–human; Sigma-Aldrich), sec16A (rabbit anti-human; Sigma-Aldrich), calreticulin (goat anti–human; Enzo Life Sciences), HA (mouse; BioLegend), TGN46 (sheep polyclonal, Bio-Rad), HA (mouse monoclonal, BioLegend; rat monoclonal BioLegend). Mounting media used in confocal and STED microscopy were either Vectashield (Vector Laboratories) or ProLong (Thermo Fisher Scientific, Waltham, Massachusetts).
Raote I., Ernst A.M., Campelo F., Rothman J.E., Pincet F, & Malhotra V. (2020). TANGO1 membrane helices create a lipid diffusion barrier at curved membranes. eLife, 9, e57822.
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4

Antibody Characterization for Cell Biology

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Antibodies used in this study were as follows: GFP (clones 7.1 and 13.1; 11814460001; Roche) (1:2,000 for Western blot (WB), 1:400 for immunofluorescence (IF)), SNX1 (clone 51/SNX1; 611482; BD) (1:1,000 for WB, 1:200 for IF), SNX2 (clone 13/SNX2; 5345661; BD) (1:1,000 for WB, 1:200 for IF), SNX6 (clone d-5,365965; Santa Cruz Biotechnology, Inc.) (1:1,000 for WB, 1:200 for IF), SNX5 (clone EPR14358; ab180520; Abcam) (WB 1:500), β-actin (A1978; Sigma-Aldrich) (1:2,000 for WB), ITGA5 (610633, BD) (1:1,000 for WB); VPS35 (97545; Abcam) (IF 1:200), GFP (GTX20290; GeneTex) (WB 1:2,000), EEA1 (C45B10; Cell Signaling Technologies) (IF 1:200), LAMP1 (clone H4A3, AB2296838; DSHB) (IF 1:200) Cathepsin D (21327-1-AP; Proteintech) (WB 1:1,000), Calnexin (ab22595; Abcam) (WB 1:1,000), TGN46 (AHP500G, Bio-Rad) (IF 1:400), Golgin-97 (clone CDF4; A-21270; Thermo Fischer) (IF 1:200), HA tag (66006-1-Ig, Proteintech) (WB 1:1,000), NCAD (clone 13A0; 14215; Cell Signaling) (WB 1:1,000), Tom20 (clone 29; 612278; BD Biosciences), GALNT2 (ab102650; Abcam) (WB 1:1,000), NRP1 (clone EPR3113; ab81321; Abcam) (WB 1:1,000, IF 1:200), NRP1 b1b2 domain (23 (link), 24 (link)) (clone 3E7.1) (IF 1:200), Tubulin (ab6046; Abcam) (WB 1:2,000), SARS-CoV-2 spike (40592-T62, Sino Biologicals) (WB 1:1,000) SARS Nucleocapsid (200-401-A50, Rockland) (IF 1:2,000).
Simonetti B., Daly J.L., Simón-Gracia L., Klein K., Weeratunga S., Antón-Plágaro C., Tobi A., Hodgson L., Lewis P.A., Heesom K.J., Shoemark D.K., Davidson A.D., Collins B.M., Teesalu T., Yamauchi Y, & Cullen P.J. (2022). ESCPE-1 mediates retrograde endosomal sorting of the SARS-CoV-2 host factor Neuropilin-1. Proceedings of the National Academy of Sciences of the United States of America, 119(25), e2201980119.
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5

Comprehensive Protein Immunodetection in Cell Biology

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The primary antibodies were used as follows: GAPDH (Proteintech, 60004-1-Ig), CapZβ (Proteintech, 25043-1-AP), ARPC1B (Sigma-Aldrich, HPA004832), p16-Arc (Abcam, ab51243), N-WASP (Novus Biologicals, NBP1-82512), His-tag (ProteinTech, 66005-1-Ig), β-actin (ProteinTech, 60008-1-Ig), α-tublin (ProteinTech, 66031-1-Ig), VPS8 (ProteinTech, 15079-1-AP), λ-tublin (Sigma-Aldrich, T6557), HSC70 (Santa Cruz, sc-24), VPS34 (CST, 4263), CapZa1/a2 (Developmental Studies Hybridoma Bank, mAb 5B12.3), anti-pan actin (Cytoskeleton, AAN02-s), RAB5A (CST, 3547 or 46449), Calnexin (ProteinTech, 10427-2-AP), LAMP1 (CST, 9091), TGN46 (Bio-Rad, AHP500GT), EEA1 (CST, 3288), MYC-tag (ProteinTech, 67447-1-Ig), Flag-tag (Sigma-Aldrich, F3165), HA-tag (Sigma-Aldrich,11867423001), StrepMAB-Classic HRP conjugate (IBA, 2-1509-001), Rabex-5 (Santa Cruz, sc-166049), Rabaptin-5 (ProteinTech, 14350-1-AP), EGFR (Santa Cruz, sc-373746), goat anti-mouse IgG (H+L) secondary antibody (Invitrogen, 31430), goat anti-rabbit IgG (H+L) secondary antibody (Invitrogen, 31460), and goat anti-rat IgG (H+L) secondary antibody (Invitrogen, 31470).
Wang D., Ye Z., Wei W., Yu J., Huang L., Zhang H, & Yue J. (2021). Capping protein regulates endosomal trafficking by controlling F-actin density around endocytic vesicles and recruiting RAB5 effectors. eLife, 10, e65910.
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6

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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7

Comprehensive Antibody Reagents for Cell Biology

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The following primary antibodies were used. Monoclonal antibodies against β-actin and GFP (Sigma-Aldrich); Shiga toxin B and ubiquitin (ThermoFisher); GM130, Gos28, Golgin-245, and syntaxin 6 (BD Biosciences, Franklin Lanes, NJ); α-tubulin (Developmental Studies Hybridoma Bank, University of Iowa); Arl1 (Abcam); VSV-G extracellular domain (David Sheff, University of Iowa Carver College of Medicine). Polyclonal antibodies against GCC88, Golgin-160, CI-M6PR, GRASP55, and GRASP65 (Proteintech); GCC185 (Abcam); GM130 (“N73,” from J. Seemann, University of Texas Southwestern Medical Center); TGN46 (Bio-Rad); LC3B (Cell signaling). Secondary antibodies were purchased from Jackson Laboratory (Bar Harbor, ME). Secondary antibodies used for fluorescence microscopy include fluorescence-labeled goat anti-mouse, goat anti-rabbit, and goat anti-sheep antibodies. Secondary antibodies used for Western blot include HRP-conjugated goat anti-mouse and goat anti-rabbit antibodies.
Ireland S.C., Huang H., Zhang J., Li J, & Wang Y. (2020). Hydrogen peroxide induces Arl1 degradation and impairs Golgi-mediated trafficking. Molecular Biology of the Cell, 31(17), 1931-1942.
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8

Comprehensive Reagents and Antibodies for Cell Biology

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All reagents were purchased from Sigma-­Aldrich, Invitrogen, Roche, Calbiochem, and Fisher unless otherwise stated. Antibodies used in this study include monoclonal antibodies against LAMP1 (H4A3; Developmental Studies Hybridoma Bank [DSHB]), LAMP2 (H4B4; DSHB), integrin β1 (P5D2; DSHB), integrin α-5 (BIIG2; DSHB), Shiga toxin B (A42L; Thermo-Fisher), and β-actin (Sigma); polyclonal antibodies against GRASP55 (ProteinTech), GRASP65 (UT465; Joachim Seemann, UT Southwestern), GM130 (N73; Joachim Seemann, UT Southwestern), Golgin-45 (ProteinTech), TGN46 (Bio-Rad), and GFP (Santa Cruz). Vesicular stomatitis virus G protein (VSV-G)-GFP adenovirus was a gift from David Sheff (University of Iowa) and Heike Fölsch (Northwestern University). The pUC19 plasmid was a gift from Daniel Klionsky (University of Michigan). pSpCas9(BB)-2A-GFP(PX458), pSpCas9(BB)-2A-Puro (PX459) plasmids are from Add­gene. Other materials used in this study include the following: tetramethylrhodamine (TRITC)-WGA (EY laboratories), TRITC-MAA (EY laboratories), fluorescein isothiocyanate (FITC)-cholera toxin B subunit (C1655, Sigma), Shiga toxin type 1 Subunit B (NR-860, BEI Resources), and puromycin (Thermo-Fisher).
Bekier ME I.I., Wang L., Li J., Huang H., Tang D., Zhang X, & Wang Y. (2017). Knockout of the Golgi stacking proteins GRASP55 and GRASP65 impairs Golgi structure and function. Molecular Biology of the Cell, 28(21), 2833-2842.
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9

Immunofluorescence Staining Protocol

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We used the following antibodies: mouse clone 9E10 against the c-Myc epitope (Covance, Princeton, NJ), mouse clone BA3R against β-ACTIN (ThermoFisher), rabbit polyclonal against Cytoskeleton-associated protein 4 (P63; Merck, Germany; cat # HPA001225), and sheep polyclonal against Trans-Golgi network integral membrane protein 2 (TGN46; Bio-Rad Laboratories, Hercules, CA; cat # AHP500G). The following fluorochrome-conjugated antibodies were from ThermoFisher: Alexa-Fluor-488-conjugated donkey antimouse IgG, Alexa-Fluor-594-conjugated donkey antirabbit IgG and Alexa-Fluor-647-conjugated donkey antisheep IgG. HRP-conjugated secondary antibodies were from Jackson ImmunoResearch (West Grove, PA). Depending on their reactivity, primary antibodies were used at a dilution 1/200 to 1/2,000. HRP-conjugated secondary antibodies were used at dilutions 1/5,000 to 1/20,000 also depending on their reactivity. All Alexa-Fluor-conjugated secondary antibodies were used at a dilution 1/1,000. The fluorescent nuclear stain 4′,6-diamidino-2-phenylindole (DAPI) was also from ThermoFisher.
Opazo J.C., Vandewege M.W., Hoffmann F.G., Zavala K., Meléndez C., Luchsinger C., Cavieres V.A., Vargas-Chacoff L., Morera F.J., Burgos P.V., Tapia-Rojas C, & Mardones G.A. (2023). How Many Sirtuin Genes Are Out There? Evolution of Sirtuin Genes in Vertebrates With a Description of a New Family Member. Molecular Biology and Evolution, 40(2), msad014.
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10

Autophagy Regulation in EV-A71 Infection

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The following antibodies were used: LC3 (Novus, NB100–2220), SQSTM1 (MBL, PM045), EGFR (Santa Cruz, SC-03), RAB5A (Cell Signaling Technology, 3547), LAMP1 (Cell Signaling Technology, 9091), TGN46 (BIORAD AHP500GT), EV-A71 (EMD Millipore, MAB979), GAPDH (EMD Millipore, MAB374), and dsRNA (English and Scientific Consulting, J2).
The following reagents were used: SsA (HKUST RDC, 10030), SsD (HKUST RDC, 10031), SsC (HKUST RDC, 10032), glycyl-L-phenyl-alanine-ß-naphthylamide (GPN, Santa Cruz, SC-252858), thapsigargin (TG, Sigma-Aldrich, T9033), bafilomycin A1 (BAF, Sigma-Aldrich, B1793), rapamycin (Sigma-Aldrich, R8781), Fura-2 AM (Invitrogen, F1221), LysoSensor Green DND-189 (Invitrogen, L7535), LysoSensor Yellow/Blue DND-160 (Invitrogen, L7545), RNAscope® Probe-V-EV-A71-PP (Advanced Cell Diagnostics, 489071), RNAscope® Multiplex Fluorescent Detection Kit (Advanced Cell Diagnostics, 320850), TFEB siRNA (L-050607-02-0005), and nontarget siRNA (Dharmacon).
Li C., Huang L., Sun W., Chen Y., He M.L., Yue J, & Ballard H. (2019). Saikosaponin D suppresses enterovirus A71 infection by inhibiting autophagy. Signal Transduction and Targeted Therapy, 4, 4.
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