- MA5-34669 - Invitrogen Antibodies DCP1A antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Dcp1a in human liver tissue lysates. Samples were incubated with Dcp1a monoclonal antibody (Product # MA5-34669), at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Dcp1a was achieved by transfecting HeLa with Dcp1a specific siRNAs (Silencer® select Product # s31548, s31549). Western blot analysis (Fig. a) was performed using whole cell extracts from the Dcp1a knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DCP1A Recombinant Rabbit Monoclonal Antibody (JB51-34) (Product # MA5-34669, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Dcp1a.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using DCP1A Recombinant Rabbit Monoclonal Antibody (JB51-34) (Product # MA5-34669) and a ~65 kDa band corresponding to Dcp1a was observed across cell lines along with an uncharacterised band (*) at ~45 kDa in the cell lines tested. Whole Cell Lysate (40 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), THP-1 (Lane 3), MOLT-4 (Lane 4), Hep G2 (Lane 5), 3T3-L1 (Lane 6), SH-SY5Y (Lane 7) and U-2 OS (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX), 12 well. 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 (1:2000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of Dcp1a 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 2% BSA for overnight at room temperature. The cells were labeled with DCP1A Recombinant Rabbit Monoclonal Antibody (JB51-34) (Product # MA5-34669) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). 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 represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Dcp1a in SiHa cells (green). Samples were fixed in paraformaldehyde and permeabilised with 0.25% Triton X100/PBS, incubated with Dcp1a monoclonal antibody (Product # MA5-34669), followed by DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Dcp1a in SH-SY-5Y cells (green). Samples were fixed in paraformaldehyde and permeabilised with 0.25% Triton X100/PBS, incubated with Dcp1a monoclonal antibody (Product # MA5-34669), followed by DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Dcp1a in LOVO cells (green). Samples were fixed in paraformaldehyde and permeabilised with 0.25% Triton X100/PBS, incubated with Dcp1a monoclonal antibody (Product # MA5-34669), followed by DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of Dcp1a in paraffin-embedded human placenta tissue. Samples were incubated with Dcp1a monoclonal antibody (Product # MA5-34669), and followed by hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of Dcp1a in paraffin-embedded human liver tissue. Samples were incubated with Dcp1a monoclonal antibody (Product # MA5-34669), and followed by hematoxylin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of Dcp1a in LOVO cells (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Samples were incubated with Dcp1a monoclonal antibody (Product # MA5-34669) at a dilution of 1:100, followed by Alexa Fluor 488-conjugated goat anti-rabbit IgG.

MA5-34669