Ubi-MCS-3FLAG-SV40-EGFP was expressed in HEK293 cells, and immunofluorescence confocal microscopy was employed to determine its subcellular localization using GM130 antibody (BD Bioscience, San Jose, CA, USA) and anti-mouse-cy3 (Jackson ImmunoResearch, West Grove, PA, USA) for the Golgi apparatus, Flag antibody (Sigma, CA, USA) and anti-rabbit-647 (Jackson ImmunoResearch) for βCGRP, and calnexin antibody (Sigma) and anti-rabbit-647 for the ER.
Gm130 antibody
Manufactured by BD
Sourced in United States
The GM130 antibody is a laboratory reagent used for the detection and localization of the GM130 protein, a Golgi matrix protein, in various cell and tissue samples. It can be utilized in techniques such as Western blotting, immunohistochemistry, and immunocytochemistry to study the cellular distribution and expression of the GM130 protein.
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Lab products found in correlation
Anti mouse cy3, by Jackson ImmunoResearch (1 mentions)
Flag antibody, by Merck Group (1 mentions)
Calnexin antibody, by Merck Group (1 mentions)
Lamp1, by Cell Signaling Technology (1 mentions)
F7425, by Merck Group (1 mentions)
Lamp2, by Santa Cruz Biotechnology (1 mentions)
Phospho sting s366, by Cell Signaling Technology (1 mentions)
Golgin 97, by Proteintech (1 mentions)
Golgin97, by Santa Cruz Biotechnology (1 mentions)
Lamp1 rabbit mab, by Cell Signaling Technology (1 mentions)
Gm130 rabbit mab, by Cell Signaling Technology (1 mentions)
Flag m2, by Merck Group (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Lsm 510 microscope, by Zeiss (1 mentions)
Vectashield dapi, by Vector Laboratories (1 mentions)
Tween 20, by Merck Group (1 mentions)
Tgn46 antibody, by Bio-Rad (1 mentions)
Ergic 53, by Enzo Life Sciences (1 mentions)
Alexa fluor secondary antibody, by Thermo Fisher Scientific (1 mentions)
β actin, by Merck Group (1 mentions)
5 protocols using gm130 antibody
1
Subcellular Localization of Ubi-MCS-3FLAG-SV40-EGFP
2
Immunoblotting and Immunofluorescence Antibody Validation
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The GOLPH3 rabbit polyclonal antibody was described previously (Dippold et al., 2009 (link)). The MYO18A antibody was a generous gift from Zissis Chroneos (University of Texas, Tyler, TX). The GM130 antibody was from BD Biosciences (San Jose, CA). The YAP antibody H-125 was from Santa Cruz Biotechnologies (Dallas, TX). The ATP synthase subunit IF1 antibody 5E2D7 used to mark mitochondria was from Invitrogen/Thermo Fisher Scientific. The LAMP1 and glyceraldehyde-3-phophate dehydrogenase (GAPDH) antibodies were from Cell Signaling Technology (Danvers, MA). The VSVG exofacial monoclonal antibody 8G5F11 was from KeraFAST (Boston, MA). The γ-tubulin antibody GTU-88 was from Sigma. The Cep-192 antibody was described previously (Wong et al., 2015 (link)). The TGN46 antibody was from Bio-Rad (Hercules, CA). The horseradish peroxidase–conjugated mouse anti-rabbit secondary antibody for Western blot was from Jackson ImmunoReasearch (West Grove, PA). Alexa Fluor (488, 594, and 647)-conjugated secondary antibodies for IF were from Invitrogen.
3
Antibody Characterization for Cellular Organelles
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Antibodies for LAMP1 (sc-20011, IF 1:200); LAMP2 (sc-18822, IF 1:200); Golgin97 (sc-59820, IF 1:500); GAPDH (sc-365062, WB 1:5000); Tubulin (sc-5286, WB 1:3000) were from Santa Cruz Biotechnology. The GM130 antibody (610822, IF 1:1000) was from BD Biosciences. Flag (M2, IF 1:1000; F7425, WB 1:3000) antibody was from Sigma. LAMP1 rabbit mAb (#9091, IF 1:200); GM130 rabbit mAb (#12480, IF 1:300); Rab5 rabbit mAb (#3547, IF 1:200), Phospho-STING S366 (#50907, WB 1:1000); Phospho-TBK1 Ser172 rabbit mAb (#5483, WB/IF 1:1000) were from Cell Signaling. The following antibodies were from Proteintech: STING (19851-1-AP, WB 1:1000, IF 1:1000); LC3 (14600-1-AP, IF 1:1000, WB 1:1000, reacts with LC3A, LC3B, and LC3C); TGN46 (13573-1-AP, IF 1:1000); Golgin97 (12640-1-AP, IF 1:1000). Alexa-488/594- and Pacific Blue-conjugated secondary antibodies were obtained from ThermoFisher Scientific.
4
Immunofluorescence Microscopy of Cultured Cells
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Cultured cells were washed twice in PBS, fixed with ice-cold 3.7% formaldehyde in PBS for 10 min, and washed with PBS. They were rinsed in PBS-0.05% Tween 20 (Sigma-Aldrich), incubated for 2 h with the primary antibodies or GM130 antibody (BD Biosciences), washed, and incubated for 1 h with the appropriate secondary antibodies conjugated to FITC or Texas Red. Coverslips were mounted in VECTASHIELD-DAPI (Vector Laboratories, Burlingame, CA), and sealed. Confocal microscopy was performed with an LSM510 microscope (Carl Zeiss, Thornwood, NY), by using excitation wavelengths of 488 nm (for FITC) and 543 nm (for Texas Red). The images were then processed and analyzed using FIJI Image J.
5
Antibody Sourcing for Cellular Imaging
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MMP-2 and Rab11 antibodies were purchased from Abcam (rabbit ab92536 and rabbit ab3612, respectively). NUCB1, β-actin, and monoclonal anti-FLAG M2 peroxidase antibodies were obtained from Sigma-Aldrich (rabbit HPA008176 and mouse A5441 and A8592, respectively); ERGIC53 was acquired from ENZO Life Sciences (mouse ENZ-ABS300); GM130 antibody and β-integrin were purchased from BD Bioscience (mouse 610822 and mouse 610467, respectively); Rab5, Rab7, and streptavidin-HRP antibodies were from Cell Signaling (rabbit C8B1, D95F2, and 3999S, respectively); and TGN46 antibody was obtained from AbD Serotec (sheep AHP500G). MMP-14 antibody was purchased from Millipore (mouse MAB3328), and GFP antibody (rabbit sc8334) and HRP-coupled secondary antibodies (anti-rabbit IgG, anti-mouse IgG, and anti-sheep IgG) were purchased from Santa Cruz Biotechnology. Sec16A antibody (KIAA0310 polyclonal rabbit) was purchased from Biomol (now Thermo Fisher Scientific, A300-648A-M). Anti-rabbit HRP antibody used with primary macrophages was purchased from Cell Signaling (7074). The Alexa Fluor secondary antibodies used for immunofluorescence (488, 594, 633, and phalloidin-Alexa Fluor 488, different species) were purchased from Thermo Fisher Scientific.
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