Human Clusterin EZ-Set™ ELISA Kit (DIY Antibody Pairs)

Clusterin (apolipoprotein J) is a 75 - 80 kDa disulfide-linked heterodimeric protein associated with the clearance of cellular debris and apoptosis.[1] In humans, clusterin is encoded by the CLU gene.This protein has several synonyms: dimeric acidic glycoprotein (DAG protein), testosterone repressed prostate message-2 (TRPM-2), sulfated glycoprotein-2 (SGP-2) and complement lysis inhibitor (CLI) . Clusterin was mapped to mouse chromosome 14. Clusterin is a ubiquitously expressed molecule thought to influence a variety of processes including cell death. In the brain, it accumulates in dying neurons following seizures and hypoxic-ischemic (H-I) injury. clusterin may be a new therapeutic target to modulate non-caspase-dependent neuronal death following acute brain injury.

Clusterin; Aging-associated gene 4 protein; Apolipoprotein J; Apo-J; Complement cytolysis inhibitor; CLI; Complement-associated protein SP-40,40; Ku70-binding protein 1; NA1/NA2; Testosterone-repressed prostate message 2; TRPM-2; Clusterin beta chain; ApoJalpha; Complement cytolysis inhibitor a chain; Clusterin alpha chain; ApoJbeta; Complement cytolysis inhibitor b chain; CLU; APOJ, CLI, KUB1; AAG4

Detected in blood plasma, cerebrospinal fluid, milk, seminal plasma and colon mucosa. Detected in the germinal center of colon lymphoid nodules and in colon parasympathetic ganglia of the Auerbach plexus (at protein level) . Ubiquitous. Detected in brain, testis, ovary, liver and pancreas, and at lower levels in kidney, heart, spleen and lung. .

Cancer, Cardiovascular, Chaperones, Complement, Immunology, Innate Immunity, Lipid Metabolism, Lipids/Lipoproteins, Metabolism, Protein Trafficking, Signal Transduction, Tumor Biomarkers

Isoform 1 functions as extracellular chaperone that prevents aggregation of nonnative proteins. Prevents stress- induced aggregation of blood plasma proteins. Inhibits formation of amyloid fibrils by APP, APOC2, B2M, CALCA, CSN3, SNCA and aggregation-prone LYZ variants (in vitro) . Does not require ATP. Maintains partially unfolded proteins in a state appropriate for subsequent refolding by other chaperones, such as HSPA8/HSC70. Does not refold proteins by itself. Binding to cell surface receptors triggers internalization of the chaperone-client complex and subsequent lysosomal or proteasomal degradation. Secreted isoform 1 protects cells against apoptosis and against cytolysis by complement. Intracellular isoforms interact with ubiquitin and SCF (SKP1-CUL1-F-box protein) E3 ubiquitin-protein ligase complexes and promote the ubiquitination and subsequent proteasomal degradation of target proteins. Promotes proteasomal degradation of COMMD1 and IKBKB. Modulates NF-kappa-B transcriptional activity. Nuclear isoforms promote apoptosis. Mitochondrial isoforms suppress BAX-dependent release of cytochrome c into the cytoplasm and inhibit apoptosis. Plays a role in the regulation of cell proliferation. .

Catalog number Description Quantity Storage of opened/reconstituted material EZ0914-CA Mouse anti-human Clusterin monoclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0914-DA Biotinylated goat anti-human Clusterin polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ0914-ST Lyophilized recombinant human Clusterin standard 50 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0914-CA Mouse anti-human Clusterin monoclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0914-DA Biotinylated goat anti-human Clusterin polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ0914-ST Lyophilized recombinant human Clusterin standard 50 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0914-CA Mouse anti-human Clusterin monoclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0914-CA Mouse anti-human Clusterin monoclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0914-DA Biotinylated goat anti-human Clusterin polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) EZ0914-DA Biotinylated goat anti-human Clusterin polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ0914-ST Lyophilized recombinant human Clusterin standard 50 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EZ0914-ST Lyophilized recombinant human Clusterin standard 50 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 O.D. 0.013 0.036 0.070 0.127 0.236 0.469 0.913 1.819 Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 O.D. 0.013 0.036 0.070 0.127 0.236 0.469 0.913 1.819 Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 Concentration (ng/ml) 0 0.78 1.56 3.12 6.25 12.5 25 50 O.D. 0.013 0.036 0.070 0.127 0.236 0.469 0.913 1.819 O.D. 0.013 0.036 0.070 0.127 0.236 0.469 0.913 1.819

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)