DDX1 Antibody (2220A) [Unconjugated]

Western blot shows lysates of Y-79 human retinoblastoma cell line, A431 human epithelial carcinoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, Neuro-2A mouse neuroblastoma cell line, and Rat-2 rat embryonic fibroblast cell line. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat DDX1 Monoclonal Antibody (Catalog # MAB9615) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). A specific band was detected for DDX1 at approximately 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

DDX1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human/Mouse/Rat DDX1 Monoclonal Antibody (Catalog # MAB9615) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

DDX1 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human/Mouse/Rat DDX1 Monoclonal Antibody (Catalog # MAB9615) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Western blot shows lysates of U2OS human osteosarcoma parental cell line and DDX1 knockout U2OS cell line (KO). Nitrocellulose membrane was probed with Rabbit Anti-Human/Mouse/Rat DDX1 Monoclonal Antibody (Catalog # MAB9615) followed by HRP-conjugated secondary antibody. A specific band was detected for DDX1 at approximately 82.4 kDa (as indicated) in the parental U2OS cell line, but is not detectable in knockout U2OS cell line. Primary antibody dilution used: 0.5 µg/ml. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

U2OS human osteosarcoma cell line lysates were prepared and immunoprecipitation was performed using 2.0 μg of Rabbit Anti-Human/Mouse/Rat DDX1 Monoclonal Antibody (Catalog # MAB9615) pre-coupled to Dynabeads Protein A. Immunoprecipitated DDX1 was detected in Western Blot with a mouse DDX1 antibody used at 1/500. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

• Reactivity: Hu, Mu, Rt
• Applications: WB, IHC, IP, ICC/IF, KO
• Clone: 2220A
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

DDX1 Antibody (2220A) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Human/mouse/rat DDX1 synthetic Peptide
  Accession # Q92499
• Specificity: Detects human, mouse, and rat DDX1 in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 1-25 ug/mL
  • Immunohistochemistry 3-25 ug/mL
  • Immunoprecipitation 2 ug/1 mg cell lysate
  • Knockout Validated
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for DDX1 Antibody (2220A) [Unconjugated]

• ATP-dependent RNA helicase DDX1
• DBP-RB
• DBP-RBDEAD box polypeptide 1
• DDX1
• DEAD (Asp-Glu-Ala-Asp) box polypeptide 1
• DEAD box protein 1
• DEAD box protein retinoblastoma
• DEAD Box-1
• DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 1
• DEAD/H-Box Helicase 1
• EC 3.6.1
• EC 3.6.1.10
• EC 3.6.4.13
• UKVH5d

Background

DEAD (Asp-Glu-Ala-Asp) box helicase 1 (DDX1) is a putative RNA helicase gene.  RNA helicases are involved in various cellular processes involving alteration of RNA secondary structure such as translation initiation, nuclear and mitochondrial splicing, and ribosome and spliceosome assembly. It has been proposed that DDX1 plays a role in RNA clearance at DNA double-strand breaks. It has also been reported as a coactivator of the NF-kappa-B promoter. DDX1 also acts as a positive transcriptional regulator of cyclin CCND2 expression. Human DDX1 has 97% homology to mouse and rat DDX1. Expression in embryos and testis suggest that DDX1 mediates cell growth and development. DDX1 has been proposed as a diagnostic marker for early recurrence in primary breast cancer.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • Q92499()