GANGLIOSIDES for Neuroscience Research

Transcription

1 GANGLIOSIDES for Neuroscience Research STANDARDS & MONOCLONAL ANTIBODIES

2 Gangliosides GANGLIOSIDE AND GLYCOLIPID STANDARDS purified PURIFIED NATURAL GANGLIOSIDES Cat. # Description Size GANGLIOSIDE A PATH A86613H Monosialoganglioside GM3, MW1214, from Human Liver), >98% purity 1 mg AMS.NS Ganglioside GM3 from Milk 5 mg A86263H Monosialoganglioside GM2, MW1375 from Human Brain, >98% purity 1 mg A86612H Monosialoganglioside GM1, MW1537 from Human Brain, >98% purity 1 mg GANGLIOSIDE B PATH AMS.NS Ganglioside GD3 Ammonium Salt from Milk A86168H Disialoganglioside GD2 MW1649 from Human Brain, >98% purity A86181H Disialoganglioside GD1b, MW1811 from Human Brain, >98% purity A86610H Trisialoganglioside GT1b, MW2085 from Human Brain, >98% purity 5 mg 1 mg 1 mg 1 mg OTHERS A86816H Monosialoganglioside GM4, MW1009 from Human Brain, >98% purity 1 mg purified SYNTHETIC GANGLIOSIDES AND OTHER GLYCOLOPIDS Cat. # Description CAS No. Formula Size AMS.GM Ganglioside GM2 (18, 18) C 67 H 121 N 3 O mg AMS.GM Ganglioside GM3 (18, 2) C 43 H 76 N 2 O mg AMS.GM Ganglioside GM3 (18,8) C 49 H 88 N 2 O mg AMS.GM Ganglioside GM3 (18, 12) C 53 H 96 N 2 O mg AMS.GM Ganglioside GM3 (18,16) C 57 H 104 N 2 O mg AMS.GM Ganglioside GM3 (18,18) C 59 H 108 N 2 O mg AMS.GM Ganglioside GM3 (18,24) C 65 H 120 N 2 O mg AMS.GX Azido-erythro-Sphingosine, 99% purity C 18 H 35 N 3 O 2 10 mg AMS.GX AMS.GX AMS.GX AMS.GX AMS.GX Azido-erythro-Sphingosine, 99% purity (2S, 3R, 4E)-2-Azido-3-Benzoyl-erythro-Sphingosine, 99% purity (2S, 3R, 4E)-2-Azido-3-Benzoyl-erythro-Sphingosine, 99% purity (2S, 3R, 4E)-2-Azido-3-(tert-Butyldimethylsilyl)-erythro-Sphingosine, 99% purity (2S, 3R, 4E)-2-Azido-3-(tert-Butyldimethylsilyl)-erythro-Sphingosine, 99% purity C 18 H 35 N 3 O 2 C 25 H 39 N 3 O 3 C 25 H 39 N 3 O 3 C 24 H 49 N 3 O 2 Si C 24 H 49 N 3 O 2 Si 100 mg 10 mg 100 mg 10 mg 100 mg 2

3 LIST OF ANTIBODIES New Cat. # Description Clone Immunogen Isotype Size S*.Cat. # GANGLIOSIDE A PATH AMS.A2576 Anti-N-Acetyl GM2 Monoclonal Antibody MK1-16 Emulsion of N-Acetyl GM2 and Lipid A IgM, κ-chain 0.1 mg AMS.A2505 Anti-GM1 Monoclonal Antibody GMB16 Purified Ganglioside IgM, κ-chain 0.1 mg AMS.A2507 Anti-GD1a Monoclonal Antibody GMR17 Purified Ganglioside IgM, κ-chain 0.1 mg AMS.A2701 Anti-GalNAc-GD1a Monoclonal Antibody 2A3D2 Salmonella minnesota strain R595 coated IgM, κ-chain 0.1 mg with ganglioside mixture from human hepatocellular carcinoma cells, PLC/PRF/5 AMS.A2702 Anti-GT1a Monoclonal Antibody GMR11 Purified Ganglioside IgM 0.1 mg AMS.A2582 Anti-GM3 Monoclonal Antibody GMR6 Purified Ganglioside IgM, κ-chain 0.1 mg GANGLIOSIDE B PATH AMS.A2580 Anti-GD3 Monoclonal Antibody GMR19 Purified Ganglioside IgM, κ-chain 0.1 mg AMS.A2508 Anti-GD1b Monoclonal Antibody GGR12 Purified Ganglioside IgG3, κ-chain 0.1 mg AMS.A2732 Anti-GT1b Monoclonal Antibody GMR5 Purified Ganglioside IgM, κ-chain 0.1 mg AMS.A2662 Anti-GQ1b Monoclonal Antibody GMR13 Purified Ganglioside IgM, κ-chain 0.1 mg GLOBOSERIES AMS.A2506 Anti-Gb3 Monoclonal Antibody BGR23 Purified Glycolipid IgG2b, κ-chain 0.1 mg LACTOSERIES AMS.A2584 Anti-Sialyl Lewis A Monoclonal Antibody 1H4 Colo201 cell IgG3, κ-chain 0.1 mg AMS.A2509 Anti-Sialyl Lewis A Monoclonal Antibody 2D3 Colo201 cell IgM, κ-chain 0.1 mg NEOLACTO SERIES AMS.A2578 Anti-Lewis X Monoclonal Antibody Lewis X Glycolipid IgM, κ-chain 0.1 mg AMS.A2510 Anti-Lewis Y Monoclonal Antibody H18A MKN cell IgG3, κ-chain 0.1 mg AMS.A2706 Anti-SGPG(HNK-1) Monoclonal Antibody NGR50 Purified sulfoglucuronosyl paragloboside IgG2a, κ-chain 0.1 mg (SGPG) adsorbed to Salmonella minnesota ** Seikagaku's catalogue number for same clone GANGLIOSIDE A PATH GANGLIOSIDE B PATH GLOBOSERIES LACTOSERIES NEOLACTO SERIES New Cat.# AMS.A2576 AMS.A2505 AMS.A2507 AMS.A2701 AMS.A2702 AMS.A2582 AMS.A2580 AMS.A2508 AMS.A2732 AMS.A2662 AMS.A2506 AMS.A2584 AMS.A2509 AMS.A2578 AMS.A2510 AMS.A2706 TLC= Thin Layer chromatography immunostaining Application TLC ELISA IHC ICC Flow WB IP Cell Cell= Cell adhesion inhibition experiments TCI antibodies available from amsbio 3Gangliosides

4 Gangliosides DISTRIBUTION OF GLYCOLIPIDS IN RAT BRAIN Distribution of Major Gangliosides in Rat Nervous System Detected by Monoclonal Antibodies Rat Region Ganglioside GM1 GD1a GD1b GT1b GQ1b Cerebellar cortex Molecular layer Purkinje cell layer Granular layer White matter Cerebral cortex* Molecular layer (I) Ext. granular layer (II) Ext. pyramidal layer (III) Int. granular layer (IV) Int. pyramidal layer (Va) Int. pyramidal layer (Vb) Polymorphic cell layer (VI) White matter Hippocampal formation Hippocampus Alveus Stratum oriens Stratus pyramidale Stratum radiatum Stratum lacunosummoleculare Dentate gyrus Polymorphic layer Granular layer Molecular layer Spinal cord Gray matter Anterior horn Posterior horn White matter Anterior funiculus Lateral funiculus Posterior funiculus Four grades have been assigned to each ganglioside labeling density. +++ = strong, ++ = moderate, + = weak, - = negative Gangliosides GM1, GD1b, GD1a, GT1b and GQ1b were detected by five MAbs. GMB16, GMR17, GGR12, GMR5 and GMR13, respectively, with indirect immunofluorescence technique. * Kotani, M. et al.: Glycobiology, 4, (1994) 4

5 DISTRIBUTION OF GLYCOLIPIDS IN RAT BRAIN Distribution Patterns of Gangliosides During Postnatal Development of Rat Cerebellar Cortex Age Ganglioside Layer of GM1 GD1a GD1b GT1b GQ1b GM3 GD3/ GD2 O-Ac O-Ac Cerebellar LD1a GD3 LD1 Cortex 1 Day Ext. granular layer b ++ d Molecular layer Purkinje cell layer Int. granular layer c White matter Day Ext. granular layer Molecular layer Purkinje cell layer Int. granular layer White matter Day Ext. granular layer Molecular layer Purkinje cell layer Int. granular layer White matter Day Ext. granular layer Molecular layer Purkinje cell layer Int. granular layer White matter Day Molecular layer Purkinje cell layer Granular layer White matter Adult Molecular layer Purkinje cell layer Granular layer White matter a MAb GMR19 reacts with both GD3 and LD1. GD3 was detected from 1 day to adulthood, whereas LD1 was expressed at later postnatal days. b External granular layer c Internal granular layer d Four grades have been assigned to each ganglioside labeling density. +++ = strong, ++ = moderate, + = weak, - = negative 5Gangliosides

6 Gangliosides APPLICATION OF ANTIBODIES Indirect Immunofluorescence Analysis of GQ1b Expression in Postnatal Rat Cerebellum Indirect Immunofluorescence Analysis of O-Ac-disialoganglioside Expression in Postnatal Rat Cerebellum A: 1 Day B: 5 Days A: 1 Day B: 5 Days C: 10 Days D: 20 Days C: 10 Days D: 20 Days E: 30 Days F: 80 Days E: 30 Days F: 80 Days Indirect Immunofluorescence Analysis of GM1 Expression in Postnatal Rat Cerebellum A: 1 Day B: 5 Days C: 10 Days D: 20 Days E: 30 Days F: 80 Days 6

7 APPLICATION OF ANTIBODIES Immunohistochemical Localization of Gangliosides in Rat Cerebellum A: anti-gm1 B: anti-gd1a C: anti-gd1b D: anti-gt1b E: anti-gq1b F: K-B Stain Immunofluorescence Staining of Small Neurons in Primary Cultured Rat Cerebellum Immunofluorescence Analysis of Ganglioside Expression in Rat Hippocampal Formation A: anti-gm1 B: anti-gd1a C: anti-gd1b D: anti-gt1b E: anti-gq1b F: K-B Stain Anti-GD1b MAb Anti-NSE Ab Phase Contrast 7Gangliosides

8 Gangliosides REFERENCES MK1-16; Monoclonal antibody (MAb) to N-acetyl GM2 1. Miyake, M. et al.: Generation of two murine monoclonal antibodies that can discriminate N-acetyl neuraminic acid residues of GM2 ganglioside. Cancer Res., 48, (1988) 2. Miyake, M. et al.: The abnormal occurrence and the differentiation-dependent distribution of N-acetyl and N-glycolyl species of the ganglioside GM2 in human germ cell tumors. Cancer, 65, (1990) GMB16; Monoclonal antibody (MAb) to GM1 1. Kotani, M. et al.: Generation of one set of monoclonal antibodies specific for a-pathway ganglio-series gangliosides. Biochem. Biophys. Acta, 1117, (1992) analysis. J. Biochem., 114, (1993) 3. Kotani, M. et al.: Developmental changes of ganglioside expressions in postnatal rat cerebellar cortex. Brain Res., 700, Kubo, H. et al.: Differential distribution of ganglioside GM1 and sulfatide during the development of Xenopus embryos. Develop. Growth.Differ., 37, Kawashima, I. et al.: Immunocytochemical analysis of gangliosides in rat primary cerebellar cultures using specific monoclonal 6. Tai, T. et al.: Cell type-specific expression of ganglioside antigens in the central nervous system. Pure and Appl. Chem., 69, A3D2; Monoclonal antibody (MAb) to GalNAc-GD1a 1. Hiraiwa, N. et al.: Gangliosides and sialoglycoproteins carrying a rare blood group antigen determinant, Cad, associated with human cancers as detected by specific monoclonal antibodies. Cancer Res., 50, (1990) 2. Zenita, K. et al.: Northern hybridization analysis of VH gene expression in murine monoclonal antibodies directed to cancer associated ganglioside antigens having various sialic acid linkages. J. Immunology. 144, (1990) GMR11; Monoclonal antibody (MAb) to GT1a 1. Kotani, M. et al.: Generation of one set of monoclonal antibodies specific for a-pathway ganglio-series gangliosides. Biochem. Biophys. Acta, 1117, (1992) analysis. J. Biochem., 114, (1993) 3. Kotani, M. et al.: Immunohistochemical localization of minor gangliosides in the rat central nervous system. Glycobiology, 4, (1994) 4. Kotani, M. et al.: Developmental changes of ganglioside expressions in postnatal rat cerebellar cortex. Brain Res., 700, Kawashima, I. et al.: Immunocytochemical analysis of gangliosides in rat primary cerebellar cultures using specific monoclonal 6. Tai, T. et al.: Cell type-specific expression of ganglioside antigens in the central nervous system. Pure and Appl. Chem., 69, GMR17; Monoclonal antibody (MAb) to GD1a 1. Kotani, M. et al.: Generation of one set of monoclonal antibodies specific for a-pathway ganglio-series gangliosides. Biochem. Biophys. Acta, 1117, (1992) analysis. J.Biochem., 114, (1993) 3. Kotani, M. et al.: Immunohistochemical localization of minor gangliosides in the rat central nervous system. Glycobiology, 4, (1994) 4. Kotani, M. et al.: Developmental changes of ganglioside expressions in postnatal rat cerebellar cortex. Brain Res., 700, Kawashima, I. et al.: Immunocytochemical analysis of gangliosides in rat primary cerebellar cultures using specific monoclonal 6. Tai, T. et al.: Cell type-specific expression of ganglioside antigens in the central nervous system. Pure and Appl. Chem., 69,

9 REFERENCES GMR6; Monoclonal antibody (MAb) to GM3 1. Kotani, M. et al.: Generation of one set of monoclonal antibodies specific for a-pathway ganglio-series gangliosides. Biochem. Biophys. Acta, 1117, (1992) analysis. J. Biochem., 114, (1993) 3. Kotani, M. et al.: Immunohistochemical localization of minor gangliosides in the rat central nervous system. Glycobiology, 4, (1994) 4. Kotani, M. et al.: Developmental changes of ganglioside expressions in postnatal rat cerebellar cortex. Brain Res., 700, Kawashima, I. et al.: Immunocytochemical analysis of gangliosides in rat primary cerebellar cultures using specific monoclonal 6. Kamisago, S. et al.: Role of sulfatides in adhesion of heliobacter pylori to gastric cancer cells. Infection and Immunity, 64, (1996) 7. Tai, T. et al.: Cell type-specific expression of ganglioside antigens in the central nervous system. Pure and Appl. Chem., 69, GMR19; Monoclonal antibody (MAb) to GD3 1. Ozawa, H. et al.: Generation of one set of monoclonal antibodies specific for b-pathway ganglio-series gangliosides. Biochem. Biophys. Acta, 1123, (1992) analysis. J.Biochem., 114, (1993) 3. Kotani, M. et al.: Immunohistochemical localization of minor gangliosides in the rat central nervous system. Glycobiology, 4, (1994) 4. Kotani, M. et al.: Developmental changes of ganglioside expressions in postnatal rat cerebellar cortex. Brain Res., 700, Kawashima, I. et al.: Immunocytochemical analysis of gangliosides in rat primary cerebellar cultures using specific monoclonal 6. Kotani, M. and Tai, T.: An immunohistochemical technique with a series of monoclonal antibodies to gangliosides, their differential distribution in the rat cerebellum. Brain Research Protocols, 1, (1997) 7. Tai, T. et al.: Cell type-specific expression of ganglioside antigens in the central nervous system. Pure and Appl. Chem., 69, GGR12; Monoclonal antibody (MAb) to GD1b 1. Ozawa, H. et al.: Generation of one set of monoclonal antibodies specific for b-pathway ganglio-series gangliosides. Biochim. Biophys. Acta, 1123, (1992) analysis. J. Biochem., 114, (1993) 3. Kotani, M. et al.: Developmental changes of ganglioside expressions in postnatal rat cerebellar cortex. Brain Res., 700, Kawashima, I. et al.: Immunocytochemical analysis of gangliosides in rat primary cerebellar cultures using specific monoclonal 5. Kotani, M. and Tai, T.: An immunohistochemical technique with a series of monoclonal antibodies to gangliosides, their differential distribution in the rat cerebellum. Brain Research Protocols, 1, (1997) 6. Tai, T. et al.: Cell type-specific expression of ganglioside antigens in the central nervous system. Pure and Appl. Chem., 69, Kawashima, I. and Tai, T.: An immunocytochemical technique with monoclonal antibodies to glycosphingolipids in rat cerebellar cultures, influence of detergent permeabilization. Brain Research Protocols, 2, (1998) 9Gangliosides

10 Gangliosides REFERENCES GMR5; Monoclonal antibody (MAb) to GT1b 1. Ozawa, H. et al.: Generation of one set of monoclonal antibodies specific for b-pathway ganglio-series gangliosides. Biochim. Biophys. Acta, 1123, (1992) analysis. J. Biochem., 114, (1993) 3. Kotani, M. et al.: Developmental changes of ganglioside expressions in postnatal rat cerebellar cortex. Brain Res., 700, Kawashima, I. et al.: Immunocytochemical analysis of gangliosides in rat primary cerebellar cultures using specific monoclonal 5. Kotani, M. and Tai, T.: An immunohistochemical technique with a series of monoclonal antibodies to gangliosides, their differen tial distribution in the rat cerebellum. Brain Research Protocols, 1, (1997) 6. Tai, T. et al.: Cell type-specific expression of ganglioside antigens in the central nervous system. Pure and Appl. Chem., 69, GMR13; Monoclonal antibody (MAb) to GQ1b 1. Ozawa, H. et al.: Generation of one set of monoclonal antibodies specific for b-pathway ganglio-series gangliosides. Biochim. Biophys. Acta, 1123, (1992) 2. Kotani,M.et al.:developmental changes of ganglioside expressions in postnatal rat cerebellar cortex.brain Res.,700, Kawashima,I.et al.:immunocytochemical analysis of gangliosides in rat primary cerebellar cultures using specific monoclonal 4. Kotani,M.and Tai,T.:An immunohistochemical technique with a series of monoclonal antibodies to gangliosides,their differential distribution in the rat cerebellum. Brain Research Protocols, 1, (1997) 5. Tai,T.et al.:cell type-specific expression of ganglioside antigens in the central nervous system.pure and Appl.Chem.,69,1903- BGR23; Monoclonal antibody (MAb) to Gb3 1. Kotani, M. et al.: Generation of one set of murine monoclonal antibodies specific for globo-series gangliosides: Evidence for differential distribution of the glycolipids in rat small intestine. Arch. Biochem. Biophys., 310, (1994) NGR50; Monoclonal antibody (MAb) to SGPG 1. Yamwaki, M. et al.: Generation and characterization of anti-sulfoglucuronosyl parasloboside monoclonal antibody NGR50 and its immunoreactivity with peripheral nerve. J. Neurosci. Res., 44, (1996) 10

11 PATHWAYS IN GANGLIOSIDE BIOSYNTHESIS Gangliosides GM3 GM2 GM1 GD1a GT1a SM3 SM4s Lac-Cer SGPG GM4 GD3 GD2 GD1b GT1b GQ1b GM1α GD1β sialylparagloboside paragloboside Gb3 globoside Forssman antigen GT3 GT2 GT1c GQ1c GP1c a pathway b pathway c pathway SSEA-4 SSEA-3 DSGG diasialylssea-3 asialo GM2 asialo GM1 GM1b GD1 α α pathway ceramide glucose galactose glucuronic N-acetyl galactosamine N-acetyl glucosamine sialic acid sulfate 11Gangliosides

12