Anti-phospho-GRASP65 (Thr220/Thr224) Antibody, clone LX108

Golgi reassembly-stacking protein 1 (UniProt: O35254; also known as Golgi peripheral membrane protein p65, Golgi reassembly-stacking protein of 65 kDa, GRASP65) is encoded by the Gorasp1 (also known as Grasp65) gene in rat. GRASP65 is a Golgi structural protein and mediates cisternae stacking via self-aggregation as well as interactions with other Golgi membrane-docking proteins, such as GM130. Golgi fragmentation results from amyloid beta accumulation, which causes activation of Cdk5 and phosphorylation of GRASP65 resulting in Golgi fragmentation. At least eight GRASP65 phosphorylation sites have been identified. GRASP65 phosphorylation during mitosis and dephosphorylation after mitosis are required for Golgi disassembly and reassembly during the cell cycle. Golgi fragmentation in Alzheimer s disease brains accelerates amyloid precursor protein (APP) trafficking and further enhances amyloid peptide (Ab) production. Expression of non-phosphorylatable mutants of GRASP65 or the inhibition of Cdk5 is shown to rescue the Golgi structure and reduce Ab40 and Ab42 production. GRASP65 mutants are also reported to reduce Ab secretion by increasing alpha cleavage of the APP. (Ref.: Joshi G., et al. (2013). Proc. Natl. Acad. Sci. USA 111(13): E1230-E1239).

~100 kDa observed. Endogenous band may run at 72 kDa and GFP-tagged will show a band at 100 kDa Uncharacterized bands may be observed in some lysate(s).

His-tagged recombinant full-length wild type GRASP65 protein treated with mitotic cytosol.

Detect GRASP65 using this mouse monoclonal Anti-phospho-GRASP65 (Thr220/Thr224) Antibody, clone LX108, Cat. No. MABS1282. It is used for Immunocytochemistry and Western Blotting.

Research Category
Signaling

Western Blotting Analysis: A representative lot detected phospho-GRASP65 (Thr220/Thr224) in Western Blotting applications (Tang, D., et. al. (2012). Biol Open. 1(12):1204-14; Joshi, G., et. al. (2013). Proc. Natl. Acad. Sci. USA. 111(13):E1230-9).

Western Blotting Analysis: A 1:500 dilution of this antibody detected phospho-GRASP65 (Thr220/Thr224) in HeLa cells stably expressing rat GRASP65-GFP.

Immunocytochemistry Analysis: A representative lot detected phospho-GRASP65 (Thr220/Thr224) in Immunocytochemistry applications (Tang, D., et. al. (2012). Biol Open. 1(12):1204-14).

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected phospho-GRASP65 (Thr220/Thr224) in rat liver Golgi and mitotic Golgi fragments.

Clone LX108 specifically recognizes GRASP65 phosphorylated on Thr220/Thr224.

Format: Purified

Protein G purified

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Isotype testing: Identity Confirmation by Isotyping Test.

Isotyping Analysis: The identity of this monoclonal antibody is confirmed by isotyping test to be mouse IgG1 .

Concentration: Please refer to lot specific datasheet.

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