Most general reagents used in this study were sourced from Sigma-Aldrich. The siRNA oligonucleotides were purchased from Dharmacon. Primary antibodies used in this study were as follows: anti-TBC1D5 [Santa Cruz, sc-376296, dilution 1:400 or 1:1000 for immunofluorescence (IF) microscopy or western blotting (WB), respectively], anti-VPS26 (Abcam, ab23892, 1:800 IF or 1:1000 WB), anti-VPS35 [Santa Cruz, sc-374372, 1:800 IF or 1:1000 WB, or from the Seaman lab (see Seaman, 2007 (link)), 1:300 for IF], anti-CIMPR (Abcam, ab2733, 1:400 IF or 1:1000 WB), anti-Lamp1 (Santa Cruz, sc-18821, 1:500 IF or 1:1000 WB), anti-Glut1 (Abcam, ab15309, 1:400 IF), anti-GM130 (BD Transduction labs 610822, 1:500 IF), anti-Fam21 (Millipore, ABT79, 1:400 IF or 1:1000 WB), anti-Aβ (Covance, SIG-39320, 1:1000 WB), anti-sAPPβ (IBL America, 10321, 1:800 WB), anti-Rab7a:GTP (NewEast Biosciences, 26923, 1:300 IF), anti-TGN46 (Seaman lab, see Seaman, 2007 (link), dilution 1:600 IF), anti-GFP (Seaman lab, see Seaman et al., 2009 (link), 1:1000 for immunoprecipitation), anti-Snx1 (BD Transduction labs, dilution 1:400 IF or 1:1000 WB) and anti-Tubulin (Sigma-Aldrich, dilution 1:1000 WB). Secondary fluorescently labelled antibodies were purchased from Invitrogen.
Anti gm130
Manufactured by BD
Sourced in United States, Germany, United Kingdom
Anti-GM130 is a laboratory reagent used to detect the GM130 protein in cells. GM130 is a structural protein that is involved in the organization and maintenance of the Golgi apparatus within eukaryotic cells. Anti-GM130 can be used in various cell biology techniques, such as immunocytochemistry and Western blotting, to study the Golgi apparatus and its related cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti flag, by Merck Group (9 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (6 mentions)
Anti β actin, by Merck Group (6 mentions)
Anti flotillin 1, by BD (5 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (5 mentions)
Hoechst 33342, by Thermo Fisher Scientific (5 mentions)
Anti cd9, by Abcam (4 mentions)
Anti tubulin, by Merck Group (4 mentions)
Anti calnexin, by Abcam (4 mentions)
Anti lc3b, by Cell Signaling Technology (3 mentions)
Anti gapdh, by Cell Signaling Technology (3 mentions)
Anti α tubulin, by Merck Group (3 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (3 mentions)
Anti tsg101, by GeneTex (3 mentions)
Anti actin, by Merck Group (3 mentions)
Anti flag m2, by Merck Group (3 mentions)
Anti lamp1, by Abcam (3 mentions)
Anti e cadherin, by BD (3 mentions)
Anti eea1, by BD (3 mentions)
Anti ha, by Roche (3 mentions)
109 protocols using anti gm130
1
Intracellular Trafficking Regulation Assay
2
Antibody Profiling for Focal Adhesion
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Antibodies used were as follows: anti-Paxillin (RRID:AB_647289 ), anti-GM130 (RRID:AB_398141 ), anti-p130Cas (RRID:AB_397667 ) and anti-RACK1 (RRID:AB_397577 ) antibodies (BD Transduction Laboratories, New Jersey, USA), anti-IFITM3 (RRID:AB_2122095 ) (Abcam, Cambridge, UK), anti-CoxIV (RRID:AB_10694213 ), anti-FAK (RRID:AB_10694068 ), anti-pFAK Y397 (RRID:AB_2173659 ), anti-pPaxillin Y118 (RRID:AB_2174466 ), anti-Rab7 (RRID:AB_1904103 ), anti-pSrc Y416 (RRID:AB_331697 ), anti-Src (clone 36D10) (RRID:AB_10693939 ), anti-LC3B (RRID:AB_2137707 ) and anti-GAPDH (RRID:AB_10622025 ) (Cell Signaling Technologies, Danvers, MA, USA), and anti-Dctn1 (RRID:AB_10842517 ), anti-Ambra1 (RRID:AB_2636939 ) and anti-pSrc Y416 (RRID:AB_309898 ) antibodies and anti-FAK (clone 4.47)-conjugated agarose antibody (RRID:AB_310789 ) (Millipore, Billerica, MA, USA). LC3B antibody was from MBL (RRID:AB_1279144 ) (MBL International, Woburn, MA, USA). TRITC-phalloidin was purchased from Life Technologies (RRID:AB_2572408 ) (Paisley, UK). Anti-rabbit (RRID:AB_2099233 ) or mouse (RRID:AB_330924 ) peroxidase-conjugated secondary antibodies were purchased from Cell Signaling Technologies. Dasatinib was obtained from Bristol Myers Squibb (Princeton, NJ, USA) and PF562271 from Pfizer (Groton, CT, USA).
3
Immunofluorescent Labeling of Golgi and PSD-95 in Hippocampal Neurons
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Example 4
For staining GM130 and TGN38, hippocampal neurons were fixed with 4% paraformaldehyde/4% sucrose in phosphate-buffered saline (PBS) and permeabilized with 0.1% Triton X-100 in PBS. Then, the cis-Golgi network and trans-Golgi network were labeled with anti-GM130 (BD Transduction Laboratories) and anti-TGN38 (Thermo Scientific), respectively, for 1 hour at room temperature. Neurons were washed and incubated with Cy3-conjugated secondary antibody for 50 minutes at room temperature. Cells were then washed with PBS 4 times and mounted on glass microscope slides in a drop of mounting medium (Thermo) containing DABCO with coverslips and applied to confocal imaging.
For staining endogenous PSD-95, cultured hippocampal neurons grown on the coverslips were fixed with 4% paraformaldehyde/4% sucrose in PBS for 15 minutes at room temperature. Then, cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and incubated with mouse anti-PSD-95 (1:200; Thermo MA1-045) mixed with 5% normal donkey serum in PBS for 1 hour at room temperature. After cells were washed 4 times with PBS, they were incubated with Cy3-conjugated anti-mouse secondary antibody for 45 minutes at room temperature. Cells were then washed with PBS, mounted on coverslips, and applied to confocal imaging.
4
Antibody Validation for Cellular Protein Localization
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Unless stated otherwise, reagents, buffers, culture media and serum for cell cultures were purchased from Sigma-Aldrich (Milan, Italy). Custom-made rabbit polyclonal anti-NS antibody and rabbit polyclonal anti-GAPDH antibody were from Abcam (Cambridge, UK). The mouse monoclonal anti-NS antibodies were made in-house as reported before (Miranda et al., 2008 (link)). Anti-KDEL was from Enzo Life Sciences (through 3VChimica S.r.l., Italy), anti-GM130 from BD Transduction Laboratories and anti-catalase from Merk Millipore (both through SIAL S.r.l., Italy). Goat polyclonal anti-rabbit-HRP (horseradish peroxidase) and rabbit anti-mouse-HRP are from Sigma-Aldrich (Milan, Italy). Goat anti-mouse IgG-Alexa Fluor® 488 and -Alexa Fluor® 594, and goat anti-rabbit IgG-Alexa Fluor® 594 were from Life Technologies (Milan, Italy).
5
Subcellular Localization of GlyR Variants
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COS7 cells transiently expressing GlyR α1 (wt) or GlyR α1 variants were stained in permeabilized cells with primary antibodies MAb4a (Synaptic Systems, Göttingen Germany) and a polyclonal anti-calnexin antibody (1:500, Abcam, Cambridge, UK) for ER staining. The detection of ERGIC was done by the monoclonal ERGIC53 antibody (1:500, Enzo Life Science, Lörrach, Germany) and cis-Golgi stainings using a monoclonal antibody anti-GM130 (1:500, BD Transduction Laboratories, Heidelberg, Germany) together with a GlyR α1 specific antibody (Chemicon, Darmstadt, Germany). Secondary antibodies used were goat anti-mouse Cy3/Alexa488 and, goat anti–rabbit Cy3 (Dianova, Hamburg, Germany) diluted 1:500. All stainings were subjected to confocal microscopy on a DMIRE2 confocal microscope.
6
Antibody Characterization for Cell Biology
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Antibodies used in this study were: anti-HA mouse monoclonal antibody (Covance Inc., Princeton, NJ, United States), anti-HA rabbit monoclonal (Upstate Cell Signaling Solutions, Waltham, MA, United States), anti-myc rabbit polyclonal, (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, United States), anti-Golgin-97 (Invitrogen, #A21270), anti-p230 (BD Transduction Laboratories, Lexington, KY, United States #611230), anti-GM130 (BD Transduction Laboratories, #51-9001978), horseradish peroxidase (HRP)-conjugated goat anti-mouse or anti-rabbit (Jackson Laboratory, Bar Harbor, ME, United States), Goat anti-GST-HRP (Amersham Biosciences, Piscataway, NJ, United States), Alexa Fluor-conjugated secondary antibodies (Invitrogen Molecular Probes, Eugene, OR, United States).
7
Analysis of NF-κB Inhibition and Golgi Markers
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NF-κB inhibitor BAY 11–7085 (SC-202490, Santa Cruz Biotechnology, Dallas, TX, USA) and monensin (M5273, Sigma Chemical Co., St. Louis, MO, USA) were dissolved in dimethyl sulfoxide (DMSO) and diluted in fresh medium before use. Antibodies used in this study and their sources are as follows: anti-golgin-97 (ab84340; Abcam Inc., Cambridge, MA, USA); anti-E-cadherin (sc-7870), anti-p65 (sc-8008), anti-GAPDH (sc-6215), and anti-Lamin A/C (sc-6215) (Santa Cruz Biotechnology, Dallas, TX, USA); anti-IκB (#4814; Cell Signaling Technology, Beverly, MA, USA); anti-phospho-NF-κB p65 (Ser529) (GTX38622; GeneTex, Irvine, CA, USA); anti-β-actin (Millipore, Bedford, MA, USA); anti-TGN46 (AD Serotec, AHP500; Bio-Rad Laboratories, Inc., Hercules, CA, USA); and anti-GM130 (610823; BD Transduction Laboratories, Lexington, KY, USA). The rabbit polyclonal antibody against human Arl1 was produced in-house as described previously [28 (link)]. Anti-GCC185 was kindly provided by Dr. Fang-Jen S. Lee, National Taiwan University, Taiwan.
8
Antibody Detection Techniques in Prion Research
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The following mouse antibodies were used: anti-Rab7 (Sigma), anti-Tsg101 (GeneTex), anti-β-actin (Abcam), anti-GM130 (BD Transduction Laboratories) and anti-prion (SAF32, Cayman chemical; AH6, TSE Resource Center,). The following rabbit antibodies were used: anti-Hrs (Novus Biologicals), anti-TGN38 (AbD Serotec), anti-GFP (Abcam), anti-EEA1 (Cell Signaling), anti-Vps26 (a gift from Juan Bonifacino, Cell Biology Metabolism Program, NICHD, NIH, Bethesda, MD), anti-CI-M6PR (a gift from Linton Traub, Department of Cell Biology, University of Pittsburgh, PA) and anti-Alix (Bethyl Laboratories). Rat anti-LAMP1 antibody (Developmental Studies Hybridoma Bank) was used. PrPc and PrPsc were routinely detected using DyL488, Cy3 and DyL647-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories). Western blots were probed using horseradish peroxidase (HRP)-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories) and InfraRed Dye 680 and 800 secondary antibodies (Li-Cor Bioscience).
9
Immunofluorescence Imaging of Organelle Markers
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Cells were seeded on glass cover slips one-day prior to infection and fixed in 4% paraformaldehyde for 20 min. Confocal images were taken on a Zeiss 710 microscope with a × 100 objective. For digitonin-mediated permeabilisation of the plasma membrane, live cells were treated with 40 μg ml−1 digitonin for 5 min on ice prior to immunolabelling with anti-CSA1 (1:400, Kirkegaard and Perry Laboratories), anti-GM130 (1:500, BD Transduction laboratories) and anti-PDI (Protein disulfide-isomerase, 1:100, Enzo) for 30 min on ice. Cells were then washed twice in PBS and fixed in 4% paraformaldehyde. After permeabilisation in PBS, 0.1% Triton X100 and 10% horse serum, cover slips were incubated with appropriate AlexaFluor secondary antibodies (Invitrogen) and DAPI (4′,6-Diamidino-2-Phenylindole, Dihydrochloride) for 30 min before mounting onto glass slides.
10
Immunolabeling of the Golgi Apparatus
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The anti-GM130 (BD Transduction Laboratories) was used at a 1:250–1:400 dilution for immunofluorescence. Hoechst (Invitrogen) was used at 1μg/ml. The mitochondrial marker MitoTracker Red CMX (Invitrogen) was used at 100nM. The secondary donkey anti-Mouse coupled to Alexa594 (Invitrogen) was used at a 1:400 dilution.
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