- MA5-15565 - Invitrogen Antibodies DDX4 antibody

Submitted references "Mitotic Slippage" and Extranuclear DNA in Cancer Chemoresistance: A Focus on Telomeres.

Salmina K, Bojko A, Inashkina I, Staniak K, Dudkowska M, Podlesniy P, Rumnieks F, Vainshelbaum NM, Pjanova D, Sikora E, Erenpreisa J
International journal of molecular sciences 2020 Apr 16;21(8)

Nuclear Factor Y (NF-Y) Modulates Encystation in Entamoeba via Stage-Specific Expression of the NF-YB and NF-YC Subunits.

Manna D, Singh U
mBio 2019 Jun 18;10(3)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of DDX4 was performed by loading 25 µg of indicated whole cell lysates and 7 µl of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. DDX4 was detected at ~75-79 kDa using a DDX4 mouse monoclonal antibody (Product # MA5-15565, upper) at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform, followed by a Goat anti-mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 31430) at a dilution of 1:20,000 for at least 30 minutes at room temperature. GAPDH was detected using GAPDH rabbit polyclonal antibody (Product # PA1-987, lower), followed by a Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036) at a dilution of 1:2000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of DDX4 was performed by loading 20 µg of indicated whole cell lysates and 7 µl of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. DDX4 was detected at ~75-79 kDa using a DDX4 mouse monoclonal antibody (Product # MA5-15565, upper) at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform followed by a Goat anti-mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 31430) at a dilution of 1:20,000 for at least 30 minutes at room temperature. GAPDH was detected using GAPDH rabbit polyclonal antibody (Product # PA1-987, lower), followed by a Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036) at a dilution of 1:2000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of DDX4 (green) in NTERA-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes at -20°C, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a DDX4mouse monoclonal antibody (Product # MA5-15565) at a concentration of 5 µg/mL in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32731) at a dilution of 1:500 for at least 30 minutes at a room temperature in the dark (green). F-actin (red) was stained with Dylight 554 Phalloidin. Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of DDX4 (green) on Human Testis tissue with a DDX4 mouse monoclonal antibody (Product # MA5-15565, right panel) at a concentration of 5 µg/mL in blocking buffer for 1 hour at room temperature compared with negative control in the absence of the primary antibody (left panel), and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, DyLight 488 conjugate at a dilution of 1:500 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with DAPI. Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometric analysis of MSCS cells using DDX4 monoclonal antibody (Product # MA5-15565) (green) and negative control (purple).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 8 Expression of meiotic and germline proteins in MS and giant cells found by Immunofluorescence--representative images for at least three experiments: ( A ) a tetraploid cell nucleus enriched with MOS-kinase (sc-86) colocalized and juxtaposed with CYCLIN B1 (DOX-D2); ( B ) an attachment of MOS to the centrosomes and microtubules of the tripolar mitosis (DOX-D4); ( C ) MOS and alpha-TUBULIN form a monopolar spindle in the early prophase (DOX-D4); ( D ) MOS is attached to interphase centrosomes (arrow) and shows a remnant of a monopolar spindle in MS (asterisk) []; ( E ) the restituting nucleus in MS becomes poor with MOS and CYCLIN B1 (DOX-D4); ( F - H ) a giant cell in MS releasing cytoplasmic DNA shows the enrichment of the cell nucleus with DMC1 (meiotic recombinase) and REC8 (meiotic cohesin) (DOX-D19); ( I ) REC8 grains are scarcely inserted in the kinetochore chains in the MS cell (arrow) (DOX-D4); ( J ) DMC1 grains are scarcely inserted in the MS cell (DOX-D7; []); ( K ) a giant cell enriched with OCT4A in the cell nucleus (a monoclonal Ab) and OCT4B in the cytoplasm (DOX-D5); and ( L ) a giant cell enriched with the germ markers, DDX4/VASA in the cell nucleus and FRAGILIS in the cytoplasm (DOX-D7). Bars = 10 um.

MA5-15565

Antibody data