Antibodies to caveolin-1 (D46G3), clathrin HC (D3C6), Rab5 (C8B1), APPLI (D83H4), EEA1 (C45B10), Rab9A (D52G8), Rab11 (D4F5) and syntaxin-6 (C34B2) rabbit antibodies were from Cell Signaling Technology. MITO-ID Red, Golgi ID Green, ER ID Red and Lyso ID Red were purchased from Enzo Life Sciences. Anti-α3 integrin (CD49c) and APC-anti-α3 integrin (CD49c) antibody (clone ASC-1), anti-CD9 antibody and FITC anti-CD9 antibody (clone H19a), PE anti-human IGF-1R (clone 1H7/CD221), PE anti-human CD115 (clone 9-4D2-1E4), PE anti-human EGFR (AY13), anti-CD59 antibody and PE anti-CD59 antibody (p282), anti-CD276 antibody and APC anti-CD276 antibody (MIH42), anti-CCR2 antibody (K036C2) and anti-CXCR4 antibody (12G5) were from Biolegend. Anti-CXCR2 antibody and APC anti-CXCR2 (Clone 6C6) were from BD Biosciences. Anti-TIMP3 antibody was from Biorbyt Ltd. (Cambridge, United Kingdom). Alexa 555 goat anti-IgG H+L antibodies were from Life Technologies. Alexa 647 anti-P2Y11 (505214) and Alexa 647 anti-MRGX2 (477533) were from R&D Systems. Anti-human LL-37 antibody (1C12) was from Hycult Biotch. Goat anti-mouse IgG (H&L):15 nm Gold was from BBI Solutions. LL-37 was synthesized by the standard FMOC chemistry, purified by reversed phase HPLC, and the mass verified by nanospray mass spectrometry. The concentration was determined by amino acid analysis (Zhang et al., 2008 (link)).
Golgi id green
Manufactured by Enzo Life Sciences
Golgi ID Green is a fluorescent dye that specifically labels the Golgi apparatus in live cells. It allows visualization of the Golgi structure using fluorescence microscopy.
Automatically generated - may contain errors
2 protocols using golgi id green
1
Antibody Panel for Vesicular Trafficking
2
Multiparametric Analysis of Protein and Organelle Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For HC and LC protein, cells (1 x 10 6 ) were fixed using a 1:1 solution of Fixation Medium A (Invitrogen) and flow cytometry buffer (PBS with 5% BSA) for 15 min at RT and then incubated with Permeabilisation Medium B (Invitrogen) containing 1:20 goat f(ab')2 antihuman IgG Alexa Fluor conjugated to 488 (Invitrogen, excitation/emission: ~500/520 nm) and goat f(ab')2 anti-human kappa conjugated to APC (Biolegend, excitation/emission: ~650/675 nm) for 15 min, before being washed and analysed. HC and LC mRNA were investigated using a PrimeFlow RNA kit with custom probes (Affymetrix, Merck) following the manufacturer protocol. Quantitation of organelles was performed by staining with Golgi-ID Green (excitation/emission: ~450/530 nm), ER-ID red (excitation/emission: ~580/660 nm), Mito-ID red (excitation/emission: ~558/690 nm; all from Enzo Life Sciences) and MitoTracker Deep Red (Invitrogen, excitation/emisssion: ~644/665 nm) using the manufacturers' protocols. DAPI (excitation/emisssion: ~340/450 nm) or Sytox nuclear (excitation/emission: ~444/480) counter-stain were used for nuclear localization.
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