ET1705-32

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

	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.
	Fig2: ICC staining of DDX5 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-32, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig3: ICC staining of DDX5 in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-32, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

	Fig4: ICC staining of DDX5 in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1705-32, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig5: 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.
	Fig6: 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.
	Fig7: 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.

	Fig8: 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).
	Fig9: 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.
	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.

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.

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