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DDX5 Recombinant Rabbit Monoclonal Antibody [JM32-50]

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Specification

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Catalog# ET1705-32

DDX5 Recombinant Rabbit Monoclonal Antibody [JM32-50]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

DDX5 Recombinant Rabbit Monoclonal Antibody [JM32-50]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human DDX5 aa 1-50 / 614.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP

Target Molecular Weight

Predicted band size: 69 kDa

Positive Control

HeLa (Human cervical adenocarcinoma cell) cell lysate, SH-SY5Y (Human neuroblastoma cell) cell lysate, MCF7 (Human breast cancer cell) cell lysate,NIH/3T3 (Mouse fibroblast) cell lysate, PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate,SH-SY5Y, NIH/3T3,PC-12, rat brain tissue, human placenta tissue, mouse brain tissue, human colon carcinoma tissue, mouse hippocampus tissue.

Conjugation

unconjugated

Clone Number

JM32-50

RRID

AB_3070564

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB IHC-P FC IF-Cell IF-Tissue
human
 
Tested Tested
Not Recommended
Tested Tested
Published
mouse
Tested Tested
Tested Tested
Not Recommended
Tested Tested
Published
rat
Tested Tested
Tested Tested
Not Recommended
Tested Tested
Published

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

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

  • 1:500-1:5,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:50-1:2,000

Target

Function

Probable ATP-dependent RNA helicase DDX5 also known as DEAD box protein 5 or RNA helicase p68 is an enzyme that in humans is encoded by the DDX5 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 RNA-dependent ATPase, and also a proliferation-associated nuclear antigen, specifically reacting with the simian virus 40 tumor antigen. This gene consists of 13 exons, and alternatively spliced transcripts containing several intron sequences have been detected, but no isoforms encoded by these transcripts have been identified.

Background References

1. Rajgor D et al. Identification of novel nesprin-1 binding partners and cytoplasmic matrin-3 in processing bodies. Mol Biol Cell 27:3894-3902 (2016).

2. Li Z et al. The long noncoding RNA THRIL regulates TNFa expression through its interaction with hnRNPL. Proc Natl Acad Sci U S A 111:1002-7 (2014).

Sequence Similarity

Belongs to the DEAD box helicase family. DDX5/DBP2 subfamily.

Post-translational Modification

Arg-502 is dimethylated, probably to asymmetric dimethylarginine.; Sumoylated; sumoylation, promoted by PIAS1, promotes interaction with HDAC1 and transcriptional repression activity. Sumoylation also significantly increases stability, and reduces polyubiquitination.; Polyubiquitinated, leading to proteasomal degradation.; Weakly phosphorylated in the G1/S phase of the cell cycle and much more at G2/M, especially at Thr and Tyr residues.

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P17844 Human

SwissProt: Q61656 Mouse

Entrez Gene: 287765 Rat

Synonyms

ATP dependent RNA helicase DDX5 antibody

DDX 5 antibody

Ddx5 antibody

DDX5_HUMAN antibody

DEAD (Asp Glu Ala Asp) box helicase 5 antibody

DEAD (Asp Glu Ala Asp) box polypeptide 5 antibody

DEAD box 5 antibody

DEAD box protein 5 antibody

DEAD/H (Asp Glu Ala Asp/His) box polypeptide 5 (RNA helicase, 68kD) antibody

G17P1 antibody

Expand

Images

  • Western blot analysis of DDX5 on different lysates with Rabbit anti-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate<br />Lane 2: SH-SY5Y (Human neuroblastoma cell) cell lysate<br />Lane 3: MCF7 (Human breast cancer cell) cell lysate<br />Lane 4: NIH/3T3 (Mouse fibroblast) cell lysate<br />Lane 5: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br />Exposure time: 6 seconds ; ECL: K1801<br /><br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>, 1/5,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 69 kDa<br />Observed band size: 69 kDa

    Western blot analysis of DDX5 on different lysates with Rabbit anti-DDX5 antibody (ET1705-32) at 1/5,000 dilution.

    Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
    Lane 2: SH-SY5Y (Human neuroblastoma cell) cell lysate
    Lane 3: MCF7 (Human breast cancer cell) cell lysate
    Lane 4: NIH/3T3 (Mouse fibroblast) cell lysate
    Lane 5: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 6 seconds ; ECL: K1801


    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET1705-32, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 69 kDa
    Observed band size: 69 kDa

  • Western blot analysis of DDX5 on different lysates with Rabbit anti-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/500 dilution.<br /><br />Lane 1: Hela cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 69 kDa<br />Observed band size: 69 kDa<br /><br />Exposure time: 2 seconds;<br /><br />10% 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/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/500 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 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of DDX5 on different lysates with Rabbit anti-DDX5 antibody (ET1705-32) at 1/500 dilution.

    Lane 1: Hela cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 69 kDa
    Observed band size: 69 kDa

    Exposure time: 2 seconds;

    10% 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 (ET1705-32) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of NIH/3T3 cells labeling DDX5 with Rabbit anti-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/250 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-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/250 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 NIH/3T3 cells labeling DDX5 with Rabbit anti-DDX5 antibody (ET1705-32) at 1/250 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-DDX5 antibody (ET1705-32) at 1/250 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.

  • Immunocytochemistry analysis of PC-12 cells labeling DDX5 with Rabbit anti-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/250 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-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/250 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 PC-12 cells labeling DDX5 with Rabbit anti-DDX5 antibody (ET1705-32) at 1/250 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-DDX5 antibody (ET1705-32) at 1/250 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.

  • Immunocytochemistry analysis of SH-SY5Y cells labeling DDX5 with Rabbit anti-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/250 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-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/250 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 SH-SY5Y cells labeling DDX5 with Rabbit anti-DDX5 antibody (ET1705-32) at 1/250 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-DDX5 antibody (ET1705-32) at 1/250 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.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</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/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</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 brain tissue with Rabbit anti-DDX5 antibody (ET1705-32) 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 (ET1705-32) 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 placenta tissue with Rabbit anti-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/100 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/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/100 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 placenta tissue with Rabbit anti-DDX5 antibody (ET1705-32) at 1/100 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 (ET1705-32) at 1/100 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 mouse brain tissue with Rabbit anti-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</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/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</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 mouse brain tissue with Rabbit anti-DDX5 antibody (ET1705-32) 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 (ET1705-32) 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-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/400 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/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</a>) at 1/400 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-DDX5 antibody (ET1705-32) at 1/400 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 (ET1705-32) at 1/400 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 mouse hippocampus tissue with Rabbit anti-DDX5 antibody (<a href="/products/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</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/ET1705-32" style="font-weight: bold;text-decoration: underline;">ET1705-32</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 mouse hippocampus tissue with Rabbit anti-DDX5 antibody (ET1705-32) 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 (ET1705-32) 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.

Citation

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