Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [4]
- Immunohistochemistry [4]
- Flow cytometry [1]
- Other assay [1]
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- Product number
- MA5-32739 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DDX5 Recombinant Rabbit Monoclonal Antibody (JM32-50)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JM32-50
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DDX5 was performed by loading whole cell lysate in 1X LDS sample buffer with 2-ME from 5 x 10^5 mouse transformed pre-B cells transduced with or without guide RNA targeting Ddx5 (gDdx5). Samples were loaded onto a 4-12 % Bis-Tris polyacrylamide gel (Product # NP0336BOX). Proteins were transferred to nitrocellulose membrane with wet/tank transfer. Membrane was blocked in 5% milk/TBST. The membrane was probe with a recombinant monoclonal anti-DDX5 antibody (Product # MA5-32739) at a dilution of 1:1,000 in 2% milk/TBST at 4°C overnight, followed by a secondary antibody HRP-anti-mouse at a dilution of 1:5,000 at room temperature for 1 hour. Chemiluminescent detection was performed. Data courtesy of Antibody Data Exchange Program.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DDX5 in different lysates using a Monoclonal antibody (Product #MA5-32739) at a dilution of 1:1,000. Positive control: Lane1: PC-12, Lane 2: MEF, Lane 3: SH-SY5Y, Lane 4: NIH-3T3, Lane 5: Mouse brain.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of DDX5 was achieved by transfecting HeLa with DDX5 specific siRNAs (Silencer® select Product # s4408, s4009). Western blot analysis (Fig. a) was performed using whole cell extracts from the DDX5 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DDX5 Recombinant Rabbit Monoclonal Antibody (JM52-30) (Product # MA5-32739, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25µg/ml, 1:4000 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 DDX5.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-DDX5 Recombinant Rabbit Monoclonal Antibody (JM52-30) (Product # MA5-32739) and a 60 kDa band corresponding to DDX5 was observed across cell lines and tissues tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), Jurkat (Lane 2), SH-SY5Y (Lane 3), Hep G2 (Lane 4), MCF7 (Lane 5), HT-29 (Lane 6), NIH/3T3 (Lane 7) and Rat testis (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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
- Immunocytochemical analysis of DDX5 in Hela cells using a DDX5 Monoclonal antibody (Product # MA5-32739) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of DDX5 in SH-SY5Y cells using a DDX5 Monoclonal antibody (Product # MA5-32739) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of DDX5 in SW480 cells using a DDX5 Monoclonal antibody (Product # MA5-32739) as seen in green. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of DDX5 was performed using 70% confluent log phase SH-SY5Y 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 DDX5 Recombinant Rabbit Monoclonal Antibody (JM52-30) (Product # MA5-32739) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 localization to the nucleus. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemical analysis of DDX5 of paraffin-embedded Mouse brain tissue using a DDX5 Monoclonal antibody (Product #MA5-32739). Counter stained with hematoxylin.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemical analysis of DDX5 of paraffin-embedded rat brain tissue using a DDX5 Monoclonal antibody (Product #MA5-32739). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemical analysis of DDX5 of paraffin-embedded Human placenta tissue using a DDX5 Monoclonal antibody (Product #MA5-32739). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of DDX5 of paraffin-embedded Mouse brain tissue using a DDX5 Monoclonal antibody (Product #MA5-32739). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of DDX5 in SH-SY5Y cells using a DDX5 Monoclonal Antibody (Product # MA5-32739) at a dilution of 1:100, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- RNA Immunoprecipitation (RIP) assay of endogenous DDX5 protein using Anti-DDX5 Antibody: RIP assay was performed using Anti-DDX5 Rabbit Recombinant Monoclonal Antibody (Product # MA5-32739, 5 ug) on whole cell lysate from HeLa cells. Normal Rabbit IgG was used as a negative IP control. RNA purified by RiboPure™ RNA Purification Kit (Product # AM1924) was analyzed by RT-PCR using the Power SYBR® Green RNA-to-CT™ 1-Step Kit (Product # 4389986) with the primers pairs over U1 snRNA, ACTB, MALAT non-coding RNA and 5S rRNA. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.