- 14-9747-82 - Invitrogen Antibodies DES antibody

Submitted references Tragedy in a heartbeat: malfunctioning desmin causes skeletal and cardiac muscle disease.

Goldfarb LG, Dalakas MC
The Journal of clinical investigation 2009 Jul;119(7):1806-13

Desmin cytoskeleton linked to muscle mitochondrial distribution and respiratory function.

Milner DJ, Mavroidis M, Weisleder N, Capetanaki Y
The Journal of cell biology 2000 Sep 18;150(6):1283-98

Desmin is essential for the tensile strength and integrity of myofibrils but not for myogenic commitment, differentiation, and fusion of skeletal muscle.

Li Z, Mericskay M, Agbulut O, Butler-Browne G, Carlsson L, Thornell LE, Babinet C, Paulin D
The Journal of cell biology 1997 Oct 6;139(1):129-44

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Desmin Monoclonal Antibody (DE-U-10), eBioscience™ (Product # 14-9747-80) and a 55 kDa band corresponding to Desmin was observed in Mouse Skeletal Muscle, Rat Skeletal Muscle, Mouse Heart, Rat Heart, Cardiomyocytes and RD cell line but not in Mouse Kidney, iPSC and MCF7. Two isoforms at ~40, ~50 kDa were also observed in Mouse Skeletal Muscle, Cardiomyocytes and RD cells. An uncharacterized band of ~25 kDa was also observed in Mouse Skeletal Muscle and Mouse Heart. Tissue extracts (30 µg lysate) of Mouse Skeletal Muscle (Lane 1), Rat Skeletal Muscle (Lane 2), Mouse Heart (Lane 3), Rat Heart (Lane 4), Mouse Kidney (Lane 7), whole cell extracts (30 µg lysate) of iPSC (Lane 5), Cardiomyocytes (Lane 6), RD (Lane 8) and MCF7 (Lane 9) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (2 ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Desmin Monoclonal Antibody (DE-U-10), eBioscience™ (Product # 14-9747-82) and a 55 kDa band corresponding to Desmin was observed in Mouse Skeletal Muscle, Mouse Heart, Cardiomyocytes but not in iPSC and Mouse Kidney. Isoforms at ~ 40 and 50 kDa were also observed in Mouse Skeletal Muscle and Cardiomyocytes. An uncharacterized band of ~25 kDa was also observed in Mouse Skeletal Muscle and Mouse Heart. Tissue extracts (30 µg lysate) of Mouse Skeletal Muscle (Lane 1), Mouse Heart (Lane 2) and Mouse Kidney (Lane 5), whole cell extracts (30 µg lysate) of iPSC (Lane 3) and Cardiomyocytes (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (5 ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry of methanol-fixed C2C12 cells using 5 µg/mL Anti-Desmin Purified, followed by F (ab')2 Anti-Mouse IgG eFluor® 570.Nuclei are stained with DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis Desmin was performed using 70% confluent log phase RD cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Desmin Monoclonal Antibody (DE-U-10), eBioscience™ (Product 14-9747-82) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A28175) at 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 cytoskeletal localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Desmin was performed using formalin-fixed paraffin-embedded mouse colon and mouse kidney tissue sections. To expose the target protein, heat-induced epitope retrieval was performed on de-paraffinized sections using eBioscience™ IHC Antigen Retrieval Solution - High pH (10X) (Product # 00-4956-58) diluted to 1X solution in water in a decloaking chamber at 110 degree Celsius for 15 minutes. Following antigen retrieval, the sections were blocked with 2% normal goat serum in 1X PBS for 45 minutes at room temperature and then probed with Desmin Monoclonal Antibody (DE-U-10), eBioscience™ (Product # 14-9747-82) at 5 µg/mL in 0.1% normal goat serum overnight at 4 degree Celsius in a humidified chamber. Detection was performed using Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 in 0.1% normal goat serum for 45 minutes at room temperature. Nuclei were stained with DAPI (Product # D1306) and the sections were mounted using ProLong™ Glass Antifade Mountant (Product # P36984). The images were captured on EVOS™ M7000 Imaging System (Product # AMF7000) at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Desmin was performed using formalin-fixed paraffin-embedded mouse colon and mouse kidney tissue sections. To expose the target protein, heat-induced epitope retrieval was performed on de-paraffinized sections using eBioscience™ IHC Antigen Retrieval Solution - High pH (10X) (Product # 00-4956-58) diluted to 1X solution in water in a decloaking chamber at 110 degree Celsius for 15 minutes. Following antigen retrieval, the sections were blocked with 2% normal goat serum in 1X PBS for 45 minutes at room temperature and then probed with Desmin Monoclonal Antibody (DE-U-10), eBioscience™ (Product # 14-9747-82) at 5 µg/mL in 0.1% normal goat serum overnight at 4 degree Celsius in a humidified chamber. Detection was performed using Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 in 0.1% normal goat serum for 45 minutes at room temperature. Nuclei were stained with DAPI (Product # D1306) and the sections were mounted using ProLong™ Glass Antifade Mountant (Product # P36984). The images were captured on EVOS™ M7000 Imaging System (Product # AMF7000) at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of Desmin was performed using formalin-fixed paraffin-embedded mouse colon and mouse kidney tissue sections. To expose the target protein, heat-induced epitope retrieval was performed on de-paraffinized sections using eBioscience™ IHC Antigen Retrieval Solution - High pH (10X) (Product # 00-4956-58) diluted to 1X solution in water in a decloaking chamber at 110 degree Celsius for 15 minutes. Following antigen retrieval, the sections were blocked with 2% normal goat serum in 1X PBS for 45 minutes at room temperature and then probed with Desmin Monoclonal Antibody (DE-U-10), eBioscience (Product # 14-9747-82) at 5 µg/mL in 0.1% normal goat serum overnight at 4 degree Celsius in a humidified chamber. Detection was performed using Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 in 0.1% normal goat serum for 45 minutes at room temperature. Nuclei were stained with DAPI (Product # D1306) and the sections were mounted using ProLong™ Glass Antifade Mountant (Product # P36984). The images were captured on EVOS™ M7000 Imaging System (Product # AMF7000) at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry of formalin-fixed paraffin embedded human testes tissue using 5 µg/mL Anti-Desmin Purified, followed by Anti-Mouse IgG Biotin, Streptavidin HRP and DAB visualization.Nuclei are counterstained with hematoxylin.

14-9747-82

Antibody data