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DDX17 Recombinant Rabbit Monoclonal Antibody [JE35-01]

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Specification

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Catalog# HA721493

DDX17 Recombinant Rabbit Monoclonal Antibody [JE35-01]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

DDX17 Recombinant Rabbit Monoclonal Antibody [JE35-01]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human DDX17 aa 100-200.

Species Reactivity

Human, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 80 kDa

Positive Control

HEK-293 cell lysate, HeLa cell lysate, K-562 cell lysate, MCF7 cell lysate, SK-OV-3 cell lysate, human breast carcinoma tissue, human colon carcinoma tissue, rat ovary tissue, MCF7.

Conjugation

unconjugated

Clone Number

JE35-01

RRID

AB_3072609

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:2,000

  • IF-Cell

  • 1:100

  • FC

  • 1:500-1:1,000

Target

Function

Probable ATP-dependent RNA helicase DDX17 (p72) is an enzyme that in humans is encoded by the DDX17 gene. 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, which is an ATPase activated by a variety of RNA species but not by dsDNA. This protein and that encoded by DDX5 gene are more closely related to each other than to any other member of the DEAD box family. Alternative splicing of this gene generates 2 transcript variants encoding different isoforms with the longer transcript reported to also initiate translation at a non-AUG (CUG) start site.

Background References

1. Zhou HZ et al. DDX17-regulated alternative splicing that produced an oncogenic isoform of PXN-AS1 to promote HCC metastasis. Hepatology. 2022 Apr

2. Xu K et al. DDX5 and DDX17-multifaceted proteins in the regulation of tumorigenesis and tumor progression. Front Oncol. 2022 Aug

Subcellular Location

Nucleus, nucleolus, Cytoplasm, cytosol.

UNIPROT

SwissProt: Q92841 Human

Entrez Gene: 315133 Rat

Synonyms

Ddx17 antibody

DDX17_HUMAN antibody

DEAD (Asp Glu Ala Asp) box helicase 17 antibody

DEAD (Asp Glu Ala Asp) box polypeptide 17 antibody

DEAD box helicase 17 antibody

DEAD box protein 17 antibody

DEAD box protein p72 antibody

DEAD/H (Asp Glu Ala Asp/His) box polypeptide 17 antibody

P72 antibody

Probable ATP-dependent RNA helicase DDX17 antibody

Expand

Images

  • Western blot analysis of DDX17 on different lysates with Rabbit anti-DDX17 antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/1,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: K-562 cell lysate (20 µg/Lane)<br />Lane 4: MCF7 cell lysate (20 µg/Lane)<br />Lane 5: SK-OV-3 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 80 kDa<br />Observed band size: 70/75 kDa<br /><br />Exposure time: 1 minute 21 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of DDX17 on different lysates with Rabbit anti-DDX17 antibody (HA721493) at 1/1,000 dilution.

    Lane 1: HEK-293 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: K-562 cell lysate (20 µg/Lane)
    Lane 4: MCF7 cell lysate (20 µg/Lane)
    Lane 5: SK-OV-3 cell lysate (20 µg/Lane)

    Predicted band size: 80 kDa
    Observed band size: 70/75 kDa

    Exposure time: 1 minute 21 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721493) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-DDX17 antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-DDX17 antibody (HA721493) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721493) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-DDX17 antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-DDX17 antibody (HA721493) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721493) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded rat ovary tissue with Rabbit anti-DDX17 antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat ovary tissue with Rabbit anti-DDX17 antibody (HA721493) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721493) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunocytochemistry analysis of MCF7 cells labeling DDX17 with Rabbit anti-DDX17 antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-DDX17 antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of MCF7 cells labeling DDX17 with Rabbit anti-DDX17 antibody (HA721493) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-DDX17 antibody (HA721493) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of MCF7 cells labeling DDX17.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721493" style="font-weight: bold;text-decoration: underline;">HA721493</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of MCF7 cells labeling DDX17.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721493, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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