Recombinant Anti-DDX5 antibody [EPR7239] (ab126730)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7239] to DDX5
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-DDX5 antibody [EPR7239]
See all DDX5 primary antibodies -
Description
Rabbit monoclonal [EPR7239] to DDX5 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- HeLa, SH-SY5Y, A431, and HT29 cell lysates; Human bladder carcinoma and Human kidney tissues; HeLa cells. IP: Mouse brain lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7239 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab126730 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
1/10000 - 1/50000. Detects a band of approximately 70 kDa (predicted molecular weight: 69 kDa).
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IP |
1/10 - 1/100.
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IHC-P | (2) |
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
1/100 - 1/250.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/10000 - 1/50000. Detects a band of approximately 70 kDa (predicted molecular weight: 69 kDa). |
IP
1/10 - 1/100. |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
Target
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Function
RNA-dependent ATPase activity. The rate of ATP hydrolysis is highly stimulated by single-stranded RNA. May be involved in pre-mRNA splicing. -
Sequence similarities
Belongs to the DEAD box helicase family. DDX5/DBP2 subfamily.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain. -
Post-translational
modificationsArg-502 is dimethylated, probably to asymmetric dimethylarginine. -
Cellular localization
Nucleus > nucleolus. - Information by UniProt
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Database links
- Entrez Gene: 1655 Human
- Entrez Gene: 13207 Mouse
- Entrez Gene: 287765 Rat
- Omim: 180630 Human
- SwissProt: P17844 Human
- SwissProt: Q61656 Mouse
- Unigene: 279806 Human
- Unigene: 220038 Mouse
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Alternative names
- ATP dependent RNA helicase DDX5 antibody
- DDX 5 antibody
- Ddx5 antibody
see all
Images
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: DDX5 knockout HAP1 whole cell lysate (20 µg)
Lane 3: MCF7 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab126730 observed at 69 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab126730 was shown to specifically react with DDX5 in wild-type cells as signal was lost in DDX5 knockout cells. Wild-type and DDX5 knockout samples were subjected to SDS-PAGE. Ab126730 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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ab126730 staining DDX5 in the human cell line HeLa (Human cervix adenocarcinoma epithelial cell) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/200. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black) Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue).
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All lanes : Anti-DDX5 antibody [EPR7239] (ab126730) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : SH-SY5Y cell lysate
Lane 3 : A431 cell lysate
Lane 4 : HT29 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 69 kDa -
Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y (human neuroblastoma) cells labelling DDX5 (green) with purified ab126730 at 1/250. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as a nuclear counterstain.
Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX5 antibody [EPR7239] (ab126730)
ab126730, at a dilution of 1/250, staining DDX5 in paraffin-embedded Human bladder carcinoma tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Purified ab126730 at 1/20 dilution (1µg) immunoprecipitating DDX5 in Mouse brain lysate.
Lane 1 (input): Mouse brain lysate 10µg
Lane 2 (+): ab126730 + Mouse brain lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126730 in Mouse brain lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/20,000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 69 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX5 antibody [EPR7239] (ab126730)
ab126730, at a dilution of 1/250, staining DDX5 in paraffin-embedded Human kidney tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab126730, at a dilution of 1/100, staining DDX5 in HeLa cells by Immunofluorescence.
Protocols
Datasheets and documents
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Datasheet download
References (13)
ab126730 has been referenced in 13 publications.
- Xu J et al. The RNA helicase DDX5 promotes viral infection via regulating N6-methyladenosine levels on the DHX58 and NF?B transcripts to dampen antiviral innate immunity. PLoS Pathog 17:e1009530 (2021). PubMed: 33909701
- Shalini V et al. Genome-wide occupancy reveals the localization of H1T2 (H1fnt) to repeat regions and a subset of transcriptionally active chromatin domains in rat spermatids. Epigenetics Chromatin 14:3 (2021). PubMed: 33407810
- Zhao X et al. Long Noncoding RNA NHEG1 Drives ß-Catenin Transactivation and Neuroblastoma Progression through Interacting with DDX5. Mol Ther 28:946-962 (2020). PubMed: 31982037
- Spadotto V et al. PRMT1-mediated methylation of the microprocessor-associated proteins regulates microRNA biogenesis. Nucleic Acids Res 48:96-115 (2020). PubMed: 31777917
- Wang T et al. LINC01116 promotes tumor proliferation and neutrophil recruitment via DDX5-mediated regulation of IL-1ß in glioma cell. Cell Death Dis 11:302 (2020). PubMed: 32358484