DDX5 Recombinant Rabbit Monoclonal Antibody [JM32-50]
cat.: ET1705-32
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JM32-50
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 69 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human DDX5 aa 1-50 / 614.
Positive control: PC-12 cell lysate, NIH/3T3 cell lysate, Hela, SH-SY5Y, SW480, rat brain tissue, human placenta tissue, mouse brain tissue, mouse brain tissue lysate, SH-SY5Y cell lysate, human colon carcinoma tissue, mouse hippocampus tissue.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:500-1:2,000
1:50-1:200
1:50-1:200
1:50-1:2,000
1:50-1:100
Uniprot #: SwissProt: P17844 Human | Q61656 Mouse
Entrez Gene: 287765 Rat
Alternative names: ATP dependent RNA helicase DDX5 DDX 5 Ddx5 DDX5_HUMAN DEAD (Asp Glu Ala Asp) box helicase 5 DEAD (Asp Glu Ala Asp) box polypeptide 5 DEAD box 5 DEAD box protein 5 DEAD/H (Asp Glu Ala Asp/His) box polypeptide 5 (RNA helicase, 68kD) G17P1 HELR HLR1 HUMP68 P68 p68 RNA helicase Probable ATP dependent RNA helicase DDX5 Probable ATP-dependent RNA helicase DDX5 RNA helicase p68
Images
ET1705-32_1.jpg Fig1: Western blot analysis of DDX5 on different lysates with Rabbit anti-DDX5 antibody (ET1705-32) at 1/500 dilution.

Lane 1: PC-12 cell lysate
Lane 2: NIH/3T3 cell lysate

Lysates/proteins at 10 µg/Lane.

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

Exposure time: 30 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.
ET1705-32_2.jpg Fig2: Western blot analysis of DDX5 on different lysates with Rabbit anti-DDX5 antibody (ET1705-32) at 1/1,000 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: SH-SY5Y cell lysate (10 µg/Lane)

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 30 seconds;

8% 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 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ET1705-32_3.jpg Fig3: 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.
ET1705-32_4.jpg Fig4: 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.
ET1705-32_5.jpg Fig5: 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.
ET1705-32_6.jpg Fig6: 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.
ET1705-32_7.jpg Fig7: 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) 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.
ET1705-32_8.jpg Fig8: 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) 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.
ET1705-32_9.jpg Fig9: 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) 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.
ET1705-32_10.jpg Fig10: 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) 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.
ET1705-32_11.jpg Fig11: 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) 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.
ET1705-32_12.jpg Fig12: Flow cytometric analysis of DDX5 was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1705-32, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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