For standard immunofluorescence experiments, cells were grown on glass-bottomed six-well plates and fixed with 4% formaldehyde. Blocking was performed with 3% BSA in PBS with 0.1% Triton X-100. Primary and secondary antibody dilutions were performed in the same solution. We used anti–protein disulfide isomerase (2792; Abcam) to detect ER, anti-LC3 (2775S; Cell Signaling Technology) to detect autophagy, anti-LAMP1 (D2D11; Cell Signaling Technology) to detect lysosomes, anticatalase (D4P7B; Cell Signaling Technology) to detect peroxisomes, and anti-GM130 (D6B1; Cell Signaling Technology) to detect the Golgi apparatus. We used as a secondary antibody Alexa Fluor 568–conjugated secondary antibodies (Life Technologies). Nuclei were stained with 1 µg/ml Hoechst 33342 (Life Technologies) during one of the post–secondary antibody washes.
Anti gm130 d6b1
Manufactured by Cell Signaling Technology
Anti-GM130 (D6B1) is a mouse monoclonal antibody that recognizes the Golgi matrix protein GM130. GM130 is a structural component of the Golgi apparatus and is involved in the organization and maintenance of Golgi structure.
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Lab products found in correlation
Anti catalase d4p7b, by Cell Signaling Technology (1 mentions)
Anti lamp1 d2d11, by Cell Signaling Technology (1 mentions)
Anti lc3 2775, by Cell Signaling Technology (1 mentions)
Alexa fluor 568 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Mouse anti gfp, by Roche (1 mentions)
Mouse anti lamp1 d4o1s, by Cell Signaling Technology (1 mentions)
Anti β actin, by Merck Group (1 mentions)
2 protocols using anti gm130 d6b1
1
Multimarker Immunofluorescence Assay
2
Antibodies Used in Immunofluorescence and Western Blotting
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The following antibodies were used in this study: rabbit Anti-LC3B (2775S; Cell Signaling Technology), for immunofluorescence and Western blotting, Anti-SQSTM1/p62 (2C11) (ab56416; Abcam) for immunofluorescence and Western blotting, rabbit Anti-LAMP1 (D2D11) XP, (9091; Cell Signaling Technology) for immunofluorescence, mouse Anti-LAMP1 (D4O1S) (15665; Cell Signaling Technology), Anti-Atg12 (Human Specific) (2010S; Cell Signaling Technology), mouse monoclonal Anti-Rab1B (SAB1400720; Sigma-Aldrich), rabbit polyclonal Anti-Rab1A (11671-1-AP; Proteintech), rabbit monoclonal Anti-GM130 (D6B1) (12480; Cell Signaling Technology), mouse monoclonal anti–β-actin (A5441; Sigma-Aldrich), mouse monoclonal Anti-Myc/c-Myc Antibody (9E10; Santa Cruz), mouse Anti-GFP (11814460001; Roche). The HRP-conjugated secondary antibodies were purchased from Cytiva, ECL Mouse IgG, HRP-linked whole Ab from sheep (NA931V) and ECL rabbit IgG, HRP-linked whole Ab from a donkey (NA934V). All Alexa-conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories, Fluorescein (FITC) AffiniPure Goat Anti-Rabbit IgG 111-095-144, Alexa Fluor 594 AffiniPure Donkey Anti-Mouse IgG 715-585-150, and Alexa Fluor 647 AffiniPure Goat Anti-Rabbit IgG 111-605-144. All the media were purchased from Sigma-Aldrich.
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