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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [2]
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- Validations
- Western blot [2]
- Immunocytochemistry [2]
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Validation data
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- Product number
- PA3-035 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ECM29 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- PA3-035 detects Ecm29 in human and mouse samples.
- Concentration
- Conc. Not Determined
Submitted references Characterization of the Brain 26S Proteasome and its Interacting Proteins.
Characterization of mammalian Ecm29, a 26 S proteasome-associated protein that localizes to the nucleus and membrane vesicles.
Tai HC, Besche H, Goldberg AL, Schuman EM
Frontiers in molecular neuroscience 2010;3
Frontiers in molecular neuroscience 2010;3
Characterization of mammalian Ecm29, a 26 S proteasome-associated protein that localizes to the nucleus and membrane vesicles.
Gorbea C, Goellner GM, Teter K, Holmes RK, Rechsteiner M
The Journal of biological chemistry 2004 Dec 24;279(52):54849-61
The Journal of biological chemistry 2004 Dec 24;279(52):54849-61
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot of Ecm29 in HeLa cells using Product # PA3-035.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed whole cell extracts (30 µg lysate) of HeLa (Lane 1), K-562 (Lane2) and NTERA-2 cl.D1 (Lane 3). The blot was probed with Anti-Ecm29 Rabbit Polyclonal Antibody (Product # PA3-035, 1:250 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 204 kDa band corresponding to Ecm29 was observed across the cell lines tested. Additional bands of 70 and 160 kDa observed, might correspond to interacting partners of dynamin intermediate chain and kinesin intermediate chain respectively. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by overnight transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Ecm29 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Ecm29 Rabbit Polyclonal Antibody (Product # PA3-035) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent staining of Ecm29 in HeLa cells using Product # PA3-035.
