Mouse Clusterin alpha Chain Antibody

Name: Mouse Clusterin alpha Chain Antibody
Brand: R&D Systems
SKU: MAB2747

Catalog #: MAB2747 Datasheet / COA / SDS

Discontinued Product

MAB2747 has been discontinued.

View all Clusterin products.

Detection of Mouse Clusterin by Western Blot.

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Mouse Clusterin alpha Chain Antibody Summary

Species Reactivity

Mouse

Specificity

Detects mouse Clusterin alpha Chain in direct ELISAs and Western blots. In Western blots, approximately 25% cross-reactivity with recombinant human Clusterin is observed.

Source

Monoclonal Rat IgG_2A Clone # 327020

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Clusterin
Glu22-Glu448
Accession # Q06890

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.

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

See below

Immunoprecipitation

25 µg/mL

Conditioned cell culture medium spiked with Recombinant Mouse Clusterin (Catalog # 2747‑HS), see our available Western blot detection antibodies

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 Mouse Clusterin by Western Blot. Western blot shows lysates of mouse brain tissue. PVDF membrane was probed with 2 µg/mL of Rat Anti-Mouse Clusterin a Chain Monoclonal Antibody (Catalog # MAB2747) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Clusterin at approximately 80 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

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-40,40, 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). Mouse Clusterin is synthesized as a precursor that contains two coiled coil domains, two nuclear localization signals (NLS), and one heparin binding domain (3-6). 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 mouse Clusterin shares 77% and 93% amino acid sequence identity with human and rat Clusterin, respectively. 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.
Lee, K.H. et al. (1993) Biochem. Biophys. Res. Commun. 194:1175.
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