Proteintech, 21773-1-AP, ZO-1 Polyclonal antibody

Size: 20ul / 150ul The ZO-1 (21773-1-AP) by Proteintech is a Polyclonal antibody targeting ZO-1 in WB, IF/ICC, IF-Fro, FC (Intra), IP, ELISA applications with reactivity to human, mouse, rat, canine, hamster samples 21773-1-AP targets ZO-1 in WB, IHC, IF/ICC, IF-Fro, FC (Intra), IP, CoIP, RIP, ELISA applications and shows reactivity with human, mouse, rat, canine, hamster samples. Tested Applications Positive WB detected in: A549 cells, HEK-293 cells, NIH/3T3 cells, Caco-2 cells, HT-29 cells, HUVEC cells, HeLa cells, HepG2 cells, K-562 cells, MCF-7 cells, mouse brain tissue, rat brain tissue, Neuro-2a cells Positive IP detected in: HEK-293 cells Positive IF-Fro detected in: mouse brain tissue Positive IF/ICC detected in: MCF-7 cells, MDCK cells, HUVEC cells Positive FC (Intra) detected in: MCF-7 cells Recommended dilution Western Blot (WB): WB : 1:5000-1:50000 Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate Immunofluorescence (IF)-FRO: IF-FRO : 1:50-1:500 Immunofluorescence (IF)/ICC: IF/ICC : 1:100-1:2000 Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.80 ug per 10^6 cells in a 100 µl suspension Background Information What is the function of ZO-1?Zona Occludens 1 (ZO-1) is a tight junction (TJ) protein found in complexes at cell-cell contacts. The role of ZO-1 is to recruit other TJ proteins.1,2The resulting TJ complexes regulate paracellular flow, contribute to apical-basal polarity, and are part of signaling pathways for proliferation and differentiation.3This protein is useful for highlighting cell-cell contacts both in cultured cells and in tissue samples, outlining cell membranes and specific structures.Where is ZO-1 expressed?The protein is localized to the cytoplasmic membrane in most types of cells, particularly where a barrier function is essential. It is common in endothelial cells,4which line the internal surface of blood and lymph vessels, and in epithelial cells,5which form the outer barrier of organs.ZO-1 is expressed in many tissues including the intestine, kidney, liver, and skeletal muscle cells.What proteins does ZO-1 interact with?ZO-1 contains multiple distinct protein domains that allow bind to other junctional proteins at the cytoplasmic membrane. The N-terminal of ZO-1 can dimerize with other ZO proteins and bind directly to other TJ proteins including claudins, connexions, and JAMs,6-10which allows it to assemble TJ complexes. The C-terminal can interact with actin and cortactin, anchoring the TJ complexes to the cytoskeleton.8,11This suggests that ZO-1 forms a link between the outer cell-cell contacts and the inner actin.What is the molecular weight of ZO-1?ZO-1 has an apparent molecular weight of 210-225 kDa. There are two isoforms of ZO-1, α+and α-, which are produced by alternative RNA splicing and differ in the presence or absence of an internal 80-amino acid domain12,13. In some literature, the bands with a molecular weight of 250-260 kDa can also be detected for ZO-114,15,16,17.1. McNeil, E., Capaldo, C. T. & Macara, I. G. Zonula occludens-1 function in the assembly of tight junctions in Madin-Darby canine kidney epithelial cells.Mol. Biol. Cell17,1922-32 (2006).2. Kratzer, I.et al.Complexity and developmental changes in the expression pattern of claudins at the blood-CSF barrier.Histochem. Cell Biol.(2012). doi:10.1007/s00418-012-1001-93. Guillemot, L., Paschoud, S., Pulimeno, P., Foglia, A. & Citi, S. The cytoplasmic plaque of tight junctions: A scaffolding and signalling center.Biochim. Biophys. Acta - Biomembr.1778,601-613 (2008).4. Tornavaca, O.et al.ZO-1 controls endothelial adherens junctions, cell-cell tension, angiogenesis, and barrier formation.J. Cell Biol.208,821-38 (2015).5. Umeda, K.et al.Establishment and characterization of cultured epithelial cells lacking expression of ZO-1.J. Biol. Chem.279,44785-94 (2004).6. Stevenson, B. R. Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia.J. Cell Biol.103,755-766 (1986).7. Itoh, M.et al.Direct Binding of Three Tight Junction-Associated Maguks, Zo-1, Zo-2, and Zo-3, with the Cooh Termini of Claudins.J. Cell Biol.147,1351-1363 (1999).8. Fanning, A. S., Jameson, B. J., Jesaitis, L. A. & Anderson, J. M. The Tight Junction Protein ZO-1 Establishes a Link between the Transmembrane Protein Occludin and the Actin Cytoskeleton.J. Biol. Chem.273,29745-29753 (1998).9.Kausalya, P. J., Reichert, M. & Hunziker, W. Connexin45 directly binds to ZO-1 and localizes to the tight junction region in epithelial MDCK cells.FEBS Lett.505,92-96 (2001).10.Itoh, M.et al.Junctional adhesion molecule (JAM) binds to PAR-3.J. Cell Biol.154,491-498 (2001).11.Itoh, M., Nagafuchi, A., Moroi, S. & Tsukita, S. Involvement of ZO-1 in Cadherin-based Cell Adhesion through Its Direct Binding to α Catenin and Actin Filaments.J. Cell Biol.138,181-192 (1997).12. Balda MS, Anderson JM. Two classes of tight junctions are revealed by ZO-1 isoforms.Am J Physiol.264,(4 Pt 1):C918-24 (1993).13. Sheth B, Fesenko I, Collins JE, Moran B, Wild AE, Anderson JM, Fleming TP. Tight junction assembly during mouse blastocyst formation is regulated by late expression of ZO-1 alpha+ isoform.Development.124,(10):2027-37(1997).14. Lassiter R, Merchen TD, Fang X, Wang Y. Protective Role of Kynurenine 3-Monooxygenase in Allograft Rejection and Tubular Injury in Kidney Transplantation.Front Immunol.7,12:671025 (2021).15. Rempe RG, Hartz AMS, Soldner ELB, Sokola BS, Alluri SR, Abner EL, Kryscio RJ, Pekcec A, Schlichtiger J, Bauer B. Matrix Metalloproteinase-Mediated Blood-Brain Barrier Dysfunction in Epilepsy.J Neurosci.2,38(18):4301-4315 (2018).16. Lin SC, Way EL. Effects of morphine and B-endorphin on Ca2+-ATPase activity of synaptic plasma membranes.NIDA Res Monogr.75,117-20(1986).17. Wu Z, Mirza H, Teo JD, Tan KS. Strain-dependent induction of human enterocyte apoptosis by blastocystis disrupts epithelial barrier and ZO-1 organization in a caspase 3- and 9-dependent manner. Biomed Res Int. 2014, 209163(2014). Specification Tested Reactivity: human, mouse, rat, canine, hamster Cited Reactivity: human, mouse, rat, pig, rabbit, canine, chicken, bovine, goat Host / Isotype: Rabbit / IgG Class: Polyclonal Type: Antibody Immunogen: CatNo: Ag16454 Product name: Recombinant human ZO1 protein Source: e coli. -derived, PGEX-4T Tag: GST Domain: 1446-1748 aa of BC111712 Sequence: SLHIHSKGAHGEGNSVSLDFQNSLVSKPDPPPSQNKPATFRPPNREDTAQAAFYPQKSFPDKAPVNGTEQTQKTVTPAYNRFTPKPYTSSARPFERKFESPKFNHNLLPSETAHKPDLSSKTPTSPKTLVKSHSLAQPPEFDSGVETFSIHAEKPKYQINNISTVPKAIPVSPSAVEEDEDEDGHTVVATARGIFNSNGGVLSSIETGVSIIIPQGAIPEGVEQEIYFKVCRDNSILPPLDKEKGETLLSPLVMCGPHGLKFLKPVELRLPHCDPKTWQNKCLPGDPNYLVGANCVSVLIDHF Predict reactive species Full Name: tight junction protein 1 (zona occludens 1) Calculated Molecular Weight: 1748 aa, 195 kDa Observed Molecular Weight: 230 kDa GenBank Accession Number: BC111712 Gene Symbol: ZO-1 Gene ID (NCBI): 7082 RRID: AB_10733242 Conjugate: Unconjugated Form: Liquid Purification Method: Antigen affinity purification UNIPROT ID: Q07157 Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3. Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20 o C storage. 20ul sizes contain 0.1% BSA.