Human Clusterin aa 22-227 Antibody

Catalog #: AF7084 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF7084
AF7084-SP

Detection of Human Clusterin beta chain by Western Blot.

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Human Clusterin aa 22-227 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Clusterin (aa 22-227) in direct ELISAs.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Clusterin isoform 1
Asp23-Arg227
Accession # P10909

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human Clusterin beta chain by Western Blot. Western blot shows lysates of human liver tissue and human serum. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Clusterin aa 22-227 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7084) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for Clusterin Precursor at approximately 60-65 kDa and Clusterin beta chain at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

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Clusterin in Human Brain. Clusterin was detected in immersion fixed paraffin-embedded sections of human Alzheimer's brain using Sheep Anti-Human Clusterin aa 22-227 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7084) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal cell bodies and processes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Clusterin

Clusterin, also known as Apolipoprotein J, Sulfated Glycoprotein 2 (SGP-2), TRPM-2, and SP-40, is a secreted multifunctional protein that was named for its ability to induce cellular clustering. It binds a wide range of molecules and may function as a chaperone of misfolded extracellular proteins. It also participates in the control of cell proliferation, apoptosis, and carcinogenesis (1, 2). Clusterin is predominantly expressed in adult testis, ovary, adrenal gland, liver, heart, and brain and in many epithelial tissues during embryonic development (3). Human Clusterin is synthesized as a precursor that contains two coiled coil domains, three nuclear localization signals (NLS), and one heparin binding domain (4‑6). Intracellular cleavages of the precursor remove the signal peptide and generate comparably sized alpha and beta chains which are secreted as an 80 kDa N-glycosylated disulfide-linked heterodimer (7, 8). Mature human Clusterin shares 77% amino acid sequence identity with mouse and rat Clusterin. High μg/mL concentrations of Clusterin circulate predominantly as a component of high density lipoprotein particles, and these are internalized and degraded through interactions with LRP-2/Megalin (9, 10). In human, an alternately spliced 50 kDa isoform of Clusterin (nCLU) lacks the signal peptide and remains intracellular (5, 11). This molecule is neither glycosylated nor cleaved into alpha and beta chains (11). In the cytoplasm, nCLU destabilizes the actin cytoskeleton and inhibits NF kappa B activation (12, 13). Cellular exposure to ionizing radiation promotes the translocation of nCLU to the nucleus where it interacts with Ku70 and promotes apoptosis (5, 11). This function contrasts with the cytoprotective effect of secreted Clusterin (14). During colon cancer tumor progression there is a downregulation of the intracellular form and an upregulation of the glycosylated secreted form (11).

References

Carver, J.A. et al. (2003) IUBMB Life 55:661.
Shannan, B. et al. (2006) Cell Death Differ. 13:12.
French, L.E. et al. (1993) J. Cell Biol. 122:1119.
Kirszbaum, L. et al. (1989) EMBO J. 8:711.
Leskov, K.S. et al. (2003) J. Biol. Chem. 278:11590.
Pankhurst, G.J. et al. (1998) Biochemistry 37:4823.
Burkey, B.F. et al. (1991) J. Lipid. Res. 32:1039.
de Silva, H.V. et al. (1990) J. Biol. Chem. 265:14292.
Jenne, D.E. et al. (1991) J. Biol. Chem. 266:11030.
Kounnas, M.Z. et al. (1995) J. Biol. Chem. 270:13070.
Pucci, S. et al. (2004) Oncogene 23:2298.
Moretti, R. M. et al. (2007) Cancer Res. 67:10325.
Santilli, G. et al. (2003) J. Biol. Chem. 278:38214.
Trougakos, I.P. et al. (2004) Cancer Res. 64:1834.

Entrez Gene IDs

1191 (Human); 12759 (Mouse)

Alternate Names

40; 40, sulfated glycoprotein 2; Aging-associated gene 4 protein; aging-associated protein 4; APOJ; apo-J; Apolipoprotein J; CLI; CLIclusterin (complement lysis inhibitor, SP-40; CLU; Clusterin; Complement cytolysis inhibitor; complement lysis inhibitor; Complement-associated protein SP-40; Ku70-binding protein 1; KUB1SGP2; MGC24903; NA1/NA2; SGP-2; SP-40; sulfated glycoprotein 2; Testosterone-repressed prostate message 2; testosterone-repressed prostate message 2, apolipoprotein J); TRPM-2; TRPM-2TRPM2