Anti-DDX3/DDX3X Antibody Picoband™

Product Name

Anti-DDX3/DDX3X Antibody Picoband™

View all DDX3 Antibodies

SKU/Catalog Number

A00751-6

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-DDX3/DDX3X Antibody Picoband™ catalog # A00751-6. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Monkey, Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-DDX3/DDX3X Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00751-6)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the N-terminus of human DDX3/DDX3X, identical to the related mouse sequences.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00751-6 is reactive to DDX3X in Human, Monkey, Mouse, Rat

Applications

A00751-6 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

73 kDa

Calculated molecular weight

73.243kDa

Background of DDX3

ATP-dependent RNA helicase DDX3X is an enzyme that in humans is encoded by the DDX3X gene. The protein encoded by this gene is a member of the large DEAD-box protein family, that is defined by the presence of the conserved Asp-Glu-Ala-Asp (DEAD) motif, and has ATP-dependent RNA helicase activity. This protein has been reported to display a high level of RNA-independent ATPase activity, and unlike most DEAD-box helicases, the ATPase activity is thought to be stimulated by both RNA and DNA. This protein has multiple conserved domains and is thought to play roles in both the nucleus and cytoplasm. Nuclear roles include transcriptional regulation, mRNP assembly, pre-mRNA splicing, and mRNA export. In the cytoplasm, this protein is thought to be involved in translation, cellular signaling, and viral replication. Misregulation of this gene has been implicated in tumorigenesis. This gene has a paralog located in the nonrecombining region of the Y chromosome. Pseudogenes sharing similarity to both this gene and the DDX3Y paralog are found on chromosome 4 and the X chromosome.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.1-0.25 μg/ml, Human, Monkey, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human, Mouse, Rat

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of DDX3/DDX3X using anti-DDX3/DDX3X antibody (A00751-6).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human HT1080 whole cell lysates,
Lane 4: monkey COS-7 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat C6 whole cell lyates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse NIH/3T3 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-DDX3/DDX3X antigen affinity purified polyclonal antibody (Catalog # A00751-6) at 0.25 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for DDX3/DDX3X at approximately 73 kDa. The expected band size for DDX3/DDX3X is at 73 kDa.

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Figure 2. IF analysis of DDX3/DDX3X using anti-DDX3/DDX3X antibody (A00751-6).
DDX3/DDX3X was detected in an immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-DDX3/DDX3X Antibody (A00751-6) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of A549 cells using anti-ADAR1/ADAR antibody (A00751-6).
Overlay histogram showing A549 cells stained with A00751-6 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ADAR1/ADAR Antibody (A00751-6, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 4. Flow Cytometry analysis of ANA-1 cells using anti-ADAR1/ADAR antibody (A00751-6).
Overlay histogram showing ANA-1 cells stained with A00751-6 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ADAR1/ADAR Antibody (A00751-6, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 5. Flow Cytometry analysis of C6 cells using anti-ADAR1/ADAR antibody (A00751-6).
Overlay histogram showing C6 cells stained with A00751-6 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ADAR1/ADAR Antibody (A00751-6, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For DDX3X (Source: Uniprot.org, NCBI)

Alternative Names

DBXCAP-Rf; DDX14; DDX3DEAD/H box-3; DEAD (Asp-Glu-Ala-Asp) box polypeptide 3, X-linked; DEAD box protein 3, X-chromosomal; DEAD box, X isoform; DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 3; EC 3.6.1; EC 3.6.4.13; helicase like protein 2; Helicase-like protein 2; HLP2ATP-dependent RNA helicase DDX3X DDX3X CAP-Rf, DBX, DDX14, DDX3, HLP2, MRX102, MRXSSB DEAD-box helicase 3 X-linked ATP-dependent RNA helicase DDX3X|DEAD (Asp-Glu-Ala-Asp) box helicase 3, X-linked|DEAD (Asp-Glu-Ala-Asp) box polypeptide 3, X-linked|DEAD box protein 3, X-chromosomal|DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 3|DEAD/H box-3|helicase-like protein 2

For more info on DDX3X, check out the DDX3X Infographic

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In this infographic, you will see the following information for DDX3X: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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