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Anti-DDX1 Picoband™ Antibody (monoclonal, 2D4)

     
  •  - Anti-DDX1 Picoband™ Antibody (monoclonal, 2D4) ABO15012
    Figure 1. Western blot analysis of DDX1 using anti-DDX1 antibody (M03727-1).
    Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
    Lane 1: human K562 whole cell lysates,
    Lane 2: human CACO-2 whole cell lysates,
    Lane 3: human U20S whole cell lysates,
    Lane 4: human HEK293 whole cell lysates,
    Lane 5: human U87 whole cell lysates,
    Lane 6: human HELA whole cell lysates,
    Lane 7: human A549 whole cell lysates,
    Lane 8: rat kidney tissue lysates,
    Lane 9: mouse kidney tissue lysates.
    After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-DDX1 antigen affinity purified monoclonal antibody (Catalog # M03727-1) at 0.25 µg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for DDX1 at approximately 86KD. The expected band size for DDX1 is at 86KD.
    detail
  •  - Anti-DDX1 Picoband™ Antibody (monoclonal, 2D4) ABO15012
    Figure 2. IHC analysis of DDX1 using anti-DDX1 antibody (M03727-1).
    DDX1 was detected in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-DDX1 Antibody (M03727-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
    detail
  •  - Anti-DDX1 Picoband™ Antibody (monoclonal, 2D4) ABO15012
    Figure 3. IHC analysis of DDX1 using anti-DDX1 antibody (M03727-1).
    DDX1 was detected in paraffin-embedded section of human pancreatic cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-DDX1 Antibody (M03727-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
    detail
  •  - Anti-DDX1 Picoband™ Antibody (monoclonal, 2D4) ABO15012
    Figure 4. IHC analysis of DDX1 using anti-DDX1 antibody (M03727-1).
    DDX1 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-DDX1 Antibody (M03727-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
    detail
  •  - Anti-DDX1 Picoband™ Antibody (monoclonal, 2D4) ABO15012
    Figure 5. IHC analysis of DDX1 using anti-DDX1 antibody (M03727-1).
    DDX1 was detected in paraffin-embedded section of human esophageal squamous carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-DDX1 Antibody (M03727-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
    detail
  •  - Anti-DDX1 Picoband™ Antibody (monoclonal, 2D4) ABO15012
    Figure 6. Flow Cytometry analysis of MCF-7 cells using anti-DDX1 antibody (M03727-1).
    Overlay histogram showing MCF-7 cells stained with M03727-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-DDX1 Antibody (M03727-1, 1 µg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
    detail
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immuno electron microscopy
  • EIA=Enzyme Immunoassay
WB, IHC, FC
Primary Accession Q92499
Host Mouse
Isotype Mouse IgG2a
Reactivity Rat, Human, Mouse
Clonality Monoclonal
Format Lyophilized
Description Anti-DDX1 Picoband™ Antibody (monoclonal, 2D4) . Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.
Reconstitution Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Additional Information
Gene ID 1653
Other Names ATP-dependent RNA helicase DDX1, 3.6.4.13, DEAD box protein 1, DEAD box protein retinoblastoma, DBP-RB, DDX1
Calculated MW 86 kDa
Application Details Western blot, 0.1-0.25 µg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5 µg/ml, Human, Rat
Flow Cytometry, 1-3 µg/1x10^6 cells, Human
Contents Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clone Names Clone: 2D4
Immunogen E.coli-derived human DDX1 recombinant protein (Position: K562-F740).
Purification Immunogen affinity purified.
Storage Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Protein Information
Name DDX1
Function Acts as an ATP-dependent RNA helicase, able to unwind both RNA-RNA and RNA-DNA duplexes. Possesses 5' single-stranded RNA overhang nuclease activity. Possesses ATPase activity on various RNA, but not DNA polynucleotides. May play a role in RNA clearance at DNA double- strand breaks (DSBs), thereby facilitating the template-guided repair of transcriptionally active regions of the genome. Together with RELA, acts as a coactivator to enhance NF-kappa-B-mediated transcriptional activation. Acts as a positive transcriptional regulator of cyclin CCND2 expression. Binds to the cyclin CCND2 promoter region. Associates with chromatin at the NF-kappa-B promoter region via association with RELA. Binds to poly(A) RNA. May be involved in 3'-end cleavage and polyadenylation of pre-mRNAs. Component of the tRNA-splicing ligase complex required to facilitate the enzymatic turnover of catalytic subunit RTCB: together with archease (ZBTB8OS), acts by facilitating the guanylylation of RTCB, a key intermediate step in tRNA ligation (PubMed:24870230). Component of a multi-helicase-TICAM1 complex that acts as a cytoplasmic sensor of viral double-stranded RNA (dsRNA) and plays a role in the activation of a cascade of antiviral responses including the induction of pro-inflammatory cytokines via the adapter molecule TICAM1. Specifically binds (via helicase ATP-binding domain) on both short and long poly(I:C) dsRNA (By similarity).
Cellular Location Nucleus. Cytoplasm. Cytoplasmic granule. Cytoplasm, cytosol {ECO:0000250|UniProtKB:Q91VR5}. Mitochondrion {ECO:0000250|UniProtKB:Q91VR5}. Note=Localized with MBNL1, TIAL1 and YBX1 in stress granules upon stress. Localized with CSTF2 in cleavage bodies. Forms large aggregates called DDX1 bodies. Relocalized into multiple foci (IR-induced foci or IRIF) after IR treatment, a process that depends on the presence of chromosomal DNA and/or RNA-DNA duplexes. Relocalized at sites of DNA double-strand breaks (DSBs) in an ATM-dependent manner after IR treatment. Colocalized with RELA in the nucleus upon TNF-alpha induction. Enters into the nucleus in case of active transcription while it accumulates in cytosol when transcription level is low (PubMed:24608264). Colocalizes in the cytosol with DDX21, DHX36 and TICAM1. Colocalizes in the mitochondria with TICAM1 and poly(I:C) RNA ligand. The multi-helicase-TICAM1 complex may translocate to the mitochondria upon poly(I:C) stimulation (By similarity) {ECO:0000250|UniProtKB:Q91VR5, ECO:0000269|PubMed:24608264}
Tissue Location Highest levels of transcription in 2 retinoblastoma cell lines and in tissues of neuroectodermal origin including the retina, brain, and spinal cord.
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Background

ATP-dependent RNA helicase DDX1 is an enzyme that in humans is encoded by the DDX1 gene. It is mapped to 2p24.3. DEAD box proteins, characterized by the conserved motif Asp-Glu-Ala-Asp (DEAD), are putative RNA helicases. They are implicated in a number of cellular processes involving alteration of RNA secondary structure such as translation initiation, nuclear and mitochondrial splicing, and ribosome and spliceosome assembly. Based on their distribution patterns, some members of this family are believed to be involved in embryogenesis, spermatogenesis, and cellular growth and division. This gene encodes a DEAD box protein of unknown function. It shows high transcription levels in 2 retinoblastoma cell lines and in tissues of neuroectodermal origin.

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$ 370.00
Cat# ABO15012
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