Anti-Ecm29 Antibody (Ab Ecm29-3)

Proteasome-associated protein ECM29 homolog (UniProt Q5VYK3; also known as Ecm29, Extracellular mutant protein 29 homolog) is encoded by the ECM29 (also known as KIAA0368) gene (Gene ID 23392) in human. Ecm29 is a large adaptor protein found exclusively on the 26S enzyme to form a subset of 26S proteasomes (Ecm proteasomes), where it functions to link 26S proteasome to molecular motors, centrosome, endoplasmic reticulum, and endocytic vesicles. Ubiquitinated proteins containing K48- and K63-linked polyubiquitin chains accumulate in the centrosomes during M phase relative to S phase, shRNA-mediated Ecm29 knockdown is found to result in a decreased accumulation of ubiquitinated proteins in the centrosomes. Ecm29 downregulation is also reported to result in an elevated level of toll-like receptor 3 (TLR3) and enhanced TLR3 signaling as a result of impaired autophagy. Ecm29 is produced as a 1845-amino acid protein whose initiator methionine is removed posttranslationally to yiled the mature protein (a.a. 2-1845) composed almost entirely of Huntingtin-elongation factor 3-protein phosphatase 2A-Tor1 (HEAT) repeats, 27 in total (a.a. 34-1828).

~205 kDa observed. 204.3 kDa calculated. A high molecular weight Ecm29 isoform is also reported to be detected by this antiserum (Ecm29-3 Ab) in mouse brain homogenate (Gorbea, C., et al., (2004). J. Biol. Chem. 2279(52):54849-54861). Uncharacterized band(s) may appear in some lysates.

Epitope: C-terminus.

Ovalbumin-conjugated linear peptide corresponding to the C-terminal sequence of human Ecm29.

Anti-Ecm29 Antibody (Ab Ecm29-3) is an antibody against Ecm29 for use in Western Blotting, Immunocytochemistry.

Research Category
Signaling

Research Sub Category
Ubiquitin & Ubiquitin Metabolism

Western Blotting Analysis: A representative lot detected the ~205 kDa target band in HeLa cell lysate and bovine brain homogenate, as well as in various mouse tissue homogenates, including testis, brain, pancreas, liver, kidney, lung, and spleen, but not heart. A high molecular weight Ecm29 isoform was also detected at a high level in mouse brain homogenate (Gorbea, C., et al., (2004). J. Biol. Chem. 2279(52):54849-54861).
Western Blotting Analysis: A representative lot detected the presence of Ecm29 in 26 proteasome-containing fractions obtained by glycerol gradient centrifugation of HeLa cell post-mitochondrial sucrose homogenate (Gorbea, C., et al., (2004). J. Biol. Chem. 2279(52):54849-54861).
Immunocytochemistry Analysis: A representative lot detected Ecm29 cellular localization at both early and late endocytic vesicles (endosomes) by fluorescent immunocytochemistry staining of methanol-fixed, 0.2% Triton X-100-permeabilized HeLa cells (Gorbea, C., et al., (2004). J. Biol. Chem. 2279(52):54849-54861).
Note: The epitope this polyclonal antiserum (Ab Ecm29-3) targets is destroyed by paraformaldehyde fixation. A 10 min incubation with -20°C 100% methanol should be used for cell fixation prior to immuncytochemistry staining by Ab Ecm29-3.

This polyclonal antiserum (Ab Ecm29-3) stained both early and late endosomes in HeLa cells and detected the ~205 kDa target band in HeLa cell lysate and in various mouse tissue homogenates. A high molecular weight Ecm29 isoform was also detected by this antiserum in mouse brain homogenate (Gorbea, C., et al., (2004). J. Biol. Chem. 2279(52):54849-54861).

Unpurified

Rabbit polyclonal antibody serum with 0.05% sodium azide.

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Evaluated by Western Blotting in MCF-7 membrane extract.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Ecm29 in 10 µg of MCF-7 membrane extract.

Concentration: Please refer to lot specific datasheet.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.