Exosomal Marker Antibody Sampler Kit

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Exosomal Marker Antibody Sampler Kit

品牌： CST

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产品介绍

产品信息

荧光素标记

Unconjugated

抗原名称

Exosomal Marker

简单描述

The Exosomal Marker Antibody Sampler Kit provides an economical means to evaluate the presence of exosomal markers. The kit includes enough primary antibody to perform two western blot experiments for each target.

研究领域

癌症,细胞生物学,纤维化,免疫学和肿瘤学,代谢,神经科学

背景

Exosomes are small membrane-bound vesicles that in recent years have emerged as important molecules for inter-cellular communication. Exosomes are produced during both normal and pathophysiological conditions, and cancer cells have been shown to secrete exosomes in greater amounts than normal cells (reviewed in 1). The exosomal markers contained in this kit are Alix, Annexin V, ICAM-1, CD9, GM130, EpCAM, flotillin, and HSP70.Alix, a cytosolic scaffold protein, regulates many cellular processes including endocytic membrane trafficking, cell adhesion through interactions with ESCRT (endosomal sorting complex required for transport) proteins, endophilins, and CIN85 (Cbl-Interacting protein of 85 kDa) (2, 3).Annexin V is a ~30 kDa protein that binds to phospho-lipids in a calcium-dependent manner (4). All annexins contain a putative PKC binding site, but only annexin V has been identified as an inhibitor of this pathway (5).Intracellular cell adhesion molecule-1 (CD54 or ICAM-1) is a cell surface glycoprotein that belongs to the immunoglobulin superfamily (IgSF) of adhesion molecules. CD54 is expressed at low levels in diverse cell types, and is induced by cytokines (TNF-alpha, interleukin-1) and bacterial lipopolysaccharides (6). Apical localization on endothelial cells (or basolateral localization on epithelial cells) is a prerequisite for leukocyte trafficking through the endothelial (or epithelial) barrier (6).The CD9 antigen belongs to the tetraspanin family of cell surface glycoproteins. Tetraspanins interact with a variety of cell surface proteins and intracellular signaling molecules in specialized tetraspanin-enriched microdomains (TEMs), where they mediate a range of processes including adhesion, motility, membrane organization, and signal transduction (7). Additional research identified CD9 as an abudant component of exosomes, and may play a role in the fusion of these secreted membrane vesicles with recipient cells (8).GM130 is required for membrane fusion events that mediate ribbon formation during Golgi assembly (9). The Golgi apparatus functions in the modification, organization, and transport of proteins and membrane targeted to other parts of the cell, such as the plasma membrane, lysosomes, and endosomes. This regulated transport is important for appropriate protein localization, secretion, and signal transduction (reviewed in 10).Epithelial cell adhesion and activation molecule (EpCAM/CD326) is a transmembrane glycoprotein that mediates calcium-independent, hemophilic adhesions on the basolateral surface of most epithelial cells (11). One of the first tumor-associated antigens discovered, EpCAM has long been a marker of epithelial and tumor tissue. Research studies have shown that EpCAM is highly expressed in cancer cells and can be used as a biomarker for the detectionof tumor-derived exposomes (reviewed in 1, 12, 13).Flotillins belong to a famiy of lipid raft-associated integral membrane proteins that are ubiquitously expressed and located to lipid rafts on the cell plasma membrane where they support signal transduction and regulate lipid raft motility and localization (14-17). In addition to its colocalization with lipid rafts on the plasma membrane, flotillin-1 also has been found at compartments of the endocytic and autophagosomal pathways, such as recycling/ late endosomes, the Golgi complex, as well as the nucleus (18, 19).HSP70 is a molecular chaperone expressed constituitively under normal conditions to maintain protein homeostatis and is induced upon environmental stress (20). HSP70 is able to interact with unfolded proteins to prevent irreversible aggregation and catalyze the refolding of their substrates in an ATP and co-chaperone dependent manner (21). An immune response is elicited upon excretion of heat shock proteins from tumor exosomes (reviewed in 1). 1.Raposo, G. and Stoorvogel, W. (2013) J Cell Biol 200, 373-83. 2.Katoh, K. et al. (2003) J Biol Chem 278, 39104-13. 3.Sadoul, R. (2006) Biol Cell 98, 69-77. 4.Huber, R. et al. (1990) EMBO J 9, 3867-74. 5.Cardó-Vila, M. et al. (2003) Mol Cell 11, 1151-62. 6.Hopkins, A.M. et al. (2004) Adv Drug Deliv Rev 56, 763-78. 7.Hemler, M.E. (2005) Nat Rev Mol Cell Biol 6, 801-11. 8.Théry, C. et al. (1999) J Cell Biol 147, 599-610. 9.Puthenveedu, M.A. et al. (2006) Nat Cell Biol 8, 238-48. 10.Barr, F.A. and Short, B. (2003) Curr Opin Cell Biol 15, 405-13. 11.Went, P.T. et al. (2004) Hum Pathol 35, 122-8. 12.Baeuerle, P.A. and Gires, O. (2007) Br J Cancer 96, 417-23. 13.Armstrong, A. and Eck, S.L. Cancer Biol Ther 2, 320-6. 14.Langhorst, M.F. et al. (2005) Cell Mol Life Sci 62, 2228-40. 15.Stuermer, C.A. and Plattner, H. (2005) Biochem Soc Symp , 109-18. 16.Fernow, I. et al. (2007) Eur J Cell Biol 86, 345-52. 17.Neumann-Giesen, C. et al. (2007) J Cell Sci 120, 395-406. 18.Liu, J. et al. (2005) J Biol Chem 280, 16125-34. 19.Santamaría, A. et al. (2005) Mol Cell Biol 25, 1900-11. 20.Nollen, E.A. and Morimoto, R.I. (2002) J Cell Sci 115, 2809-16. 21.Young, J.C. et al. (2003) Trends Biochem Sci 28, 541-7.

研究领域

癌症,细胞生物学,纤维化,免疫学和肿瘤学,代谢,神经科学

翻译后修饰

unmodified

数据库链接

Entrez-Gene ID

3383,2801,4072,10211,928,308,3303,10015

UniProt ID

P05362,Q08379,P16422,O75955,P21926,P08758,P0DMV8,Q8WUM4

参考图片

Immunohistochemical analysis of paraffin-embedded Ramos cell pellet (left, positive) or A549 cell pellet (right, negative) using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb.

Western blot analysis of extracts from various cell lines using GM130 (D6B1) XP® Rabbit mAb.

Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using GM130 (D6B1) XP® Rabbit mAb #12480. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Confocal immunofluorescent analysis of human iPSC-derived cortical glutamatergic neurons at 7 days in vitro using GM130 (D6B1) XP® Rabbit mAb (green), β3-Tubulin (E9F3E) Mouse mAb #45058 (red), and DAPI #4083 (blue). iCell GlutaNeurons were kindly provided by FUJIFILM Cellular Dynamics, Inc.

Confocal immunofluorescent analysis of human iPSC-derived astrocyte cells at 7 days in vitro using GM130 (D6B1) XP® Rabbit mAb (green), GFAP (GA5) Mouse mAb #3670 (red), and DAPI #4083 (blue). iCell Astrocytes 2.0 were kindly provided by FUJIFILM Cellular Dynamics, Inc.

Western blot analysis of extracts from various cell lines using CD9 (D8O1A) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Simple Western™ analysis of lysates (1.0 mg/mL) from HCT-116 cells using CD9 (D8O1A) Rabbit mAb #13174. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Western blot analysis of extracts from various cell lines using Flotillin-1 (D2V7J) XP® Rabbit mAb.

Western blot analysis of cell extracts from various cell types using Alix (3A9) Mouse mAb.

Immunoprecipitation of Alix from HeLa cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is Alix (3A9) Mouse mAb, #2171. Western blot was performed using Alix (3A9) Mouse mAb.

Western blot analysis of extracts from MCF7 (EpCAM positive), HT-29 (EpCAM positive), and HeLa (EpCAM negative) cells using EpCAM (D1B3) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western blot analysis of extracts from HeLa, NIH/3T3 and C6 cells, using HSP70 (D69) Antibody.

Western blot analysis of extracts from A549 cells, untreated (-) or treated with hTNF-α #8902 (10 ng/mL, 6 hr; +), using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.

Western blot analysis of extracts from various cell lines using Annexin V Antibody.

Immunoprecipitation of GM130 protein from ZR-75-1 cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or GM130 (D6B1) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using GM130 (D6B1) XP® Rabbit mAb.

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human CD9 protein (hCD9-Myc/DDK; +), using CD9 (D8O1A) Rabbit mAb.

Immunoprecipitation of flotillin-1 protein from BT-20 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Flotillin-1 (D2V7J) XP® Rabbit mAb. Western blot analysis was performed using Flotillin-1 (D2V7J) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Brefeldin A #9972 (200 μM, 30 min; right), using GM130 (D6B1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Flotillin-1 (D2V7J) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb performed on the Leica® BOND™ Rx.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb.

Confocal immunofluorescent analysis of BT-20 cells using Flotillin-1 (D2V7J) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human tonsil using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded normal rhesus monkey spleen using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using CD54/ICAM-1 (E3Q9N) XP® Rabbit mAb (left) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).

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货号：

74220T

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