HA722377

DDX39A Recombinant Rabbit Monoclonal Antibody [PSH05-57]

cat.: HA722377

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse
Applications:	WB, IF-Cell, IHC-P, IP, IF-Tissue
Clonality:	Monoclonal
Clone number:	PSH05-57

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 49 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human DDX39A aa 1-50 / 427.
Positive control:	Jurkat cell lysate, HeLa cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, mouse testis tissue lysate, HeLa, human colon tissue, mouse colon tissue.
Subcellular location:	Nucleus, Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P IP IF-Tissue	1:1,000 1:100 1:1,000-1:5,000 1-2μg/sample 1:200-1:1,000
Uniprot #:	SwissProt: O00148 Human \| Q8VDW0 Mouse
Alternative names:	ATP dependent RNA helicase DDX39 ATP-dependent RNA helicase DDX39A BAT1 BAT1L DDX39 DDXL DEAD (Asp-Glu-Ala-Asp) box polypeptide 39 isoform 2 DEAD (Asp-Glu-Ala-Asp) box polypeptide 39A DEAD box protein 39 DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 39 Eukaryotic initiation factor 4a Nuclear RNA helicase URH49 Nuclear RNA helicase, DECD variant of DEAD box family UAP56 related helicase UAP56 related helicase, 49 kDa URH49 wu:fc87b12 zgc:55881

Images

Fig1: Western blot analysis of DDX39A on different lysates with Rabbit anti-DDX39A antibody (HA722377) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: Jurkat cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: HepG2 cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
Lane 5: Mouse testis tissue lysate (10 µg/Lane)

Predicted band size: 49 kDa
Observed band size: 49 kDa

Exposure time: 25 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 (HA722377) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of DDX39A on different lysates with Rabbit anti-DDX39A antibody (HA722377) at 1/1,000 dilution.

Lane 1: RPE-si NT cell lysate
Lane 2: RPE-si DDX39A cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 49 kDa
Observed band size: 49 kDa

Exposure time: 42 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 (HA722377) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of HeLa cells labeling DDX39A with Rabbit anti-DDX39A antibody (HA722377) 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-DDX39A antibody (HA722377) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-DDX39A antibody (HA722377) at 1/5,000 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 (HA722377) at 1/5,000 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 mouse colon tissue with Rabbit anti-DDX39A antibody (HA722377) at 1/1,000 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 (HA722377) 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: DDX39A was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722377 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722377 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA722377 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA722377 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 10 seconds; ECL: K1801

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