HA500374

DDX58 Rabbit Polyclonal Antibody

cat.: HA500374

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse
Applications:	WB, IHC-P, IF-Cell, FC(Intra)
Clonality:	Polyclonal

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2.1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 107 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human DDX58 aa1-250.
Positive control:	Jurkat (Human T-lymphoblastic cells) cell lysate, HEK-293 (Human embryonic kidney cell) cell lysate, HeLa (Human cervical adenocarcinoma cell) cell lysate, A549 (Human lung adenocarcinoma cell) cell lysate, RAW264.7 (Mouse monocytic macrophage leukemia cell) cell lysate, RAW264.7 treated with 1 μg/mL LPS for 24 hours cell lysate, rat bladder tissue, human kidney tissue, human stomach tissue, mouse large intestine tissue, Hela, A549.
Subcellular location:	Cytoskeleton, Plasma membrane, Cytoplasm.
Recommended Dilutions: WB IHC-P IF-Cell FC(Intra)	1:5,000 1:400-1:1,000 1:200 1:500-1:1,000
Uniprot #:	SwissProt: O95786 Human \| Q6Q899 Mouse
Alternative names:	Ddx58 DDX58_HUMAN DEAD (Asp Glu Ala Asp) box polypeptide 58 DEAD (Asp Glu Ala Asp/His) box polypeptide DEAD box protein 58 DEAD/H (Asp Glu Ala Asp/His) box polypeptide RIG1 DKFZp434J1111 DKFZp686N19181 FLJ13599 Probable ATP dependent RNA helicase DDX58 Probable ATP-dependent RNA helicase DDX58 Retinoic acid inducible gene 1 protein Retinoic acid-inducible gene 1 protein Retinoic acid-inducible gene I protein RIG I Rig-1 RIG-I RIG1 rigi RLR 1 RNA helicase RNA helicase RIG I SGMRT2

Images

Fig1: Western blot analysis of DDX58 on different lysates with Rabbit anti-DDX58 antibody (HA500374) at 1/5,000 dilution.

Lane 1: Jurkat (Human T-lymphoblastic cells) cell lysate
Lane 2: HEK-293 (Human embryonic kidney cell) cell lysate
Lane 3: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 4: A549 (Human lung adenocarcinoma cell) cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 20 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA500374, 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: 107 kDa
Observed band size: 100 kDa

Fig2: Western blot analysis of DDX58 on different lysates with Rabbit anti-DDX58 antibody (HA500374) at 1/5,000 dilution.

Lane 1: RAW264.7 (Mouse monocytic macrophage leukemia cell) cell lysate
Lane 2: RAW264.7 treated with 1 μg/mL LPS for 24 hours cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 20 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA500374, 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: 107 kDa
Observed band size: 100 kDa

	Fig3: Immunohistochemical analysis of paraffin-embedded rat bladder tissue with Rabbit anti-DDX58 antibody (HA500374) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA500374) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-DDX58 antibody (HA500374) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA500374) at 1/1,000 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-DDX58 antibody (HA500374) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA500374) at 1/1,000 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 mouse large intestine tissue with Rabbit anti-DDX58 antibody (HA500374) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA500374) 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.

Fig7: Immunocytochemistry analysis of Hela cells labeling DDX58 with Rabbit anti-DDX58 antibody (HA500374) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-DDX58 antibody (HA500374) at 1/200 dilution in 2% negative goat serum 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.

Fig8: Flow cytometric analysis of A549 cells labeling DDX58.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500374, 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).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.