Invitrogen

DDX4 Monoclonal Antibody (2F9H5)

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DDX4 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of DDX4 (green) in NTERA-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes at -20°C, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a DDX4mouse monoclonal antibody (Product # MA5-15565) at a concentration of 5 µg/mL in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32731) at a dilution of 1:500 for at least 30 minutes at a room temperature in the dark (green). F-actin (red) was stained with Dylight 554 Phalloidin. Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

DDX4 Antibody in Immunohistochemistry (IHC) — Immunofluorescent analysis of DDX4 (green) on Human Testis tissue with a DDX4 mouse monoclonal antibody (Product # MA5-15565, right panel) at a concentration of 5 µg/mL in blocking buffer for 1 hour at room temperature compared with negative control in the absence of the primary antibody (left panel), and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, DyLight 488 conjugate at a dilution of 1:500 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with DAPI. Images were taken at 60X magnification.

DDX4 Antibody in Western Blot (WB) — Western blot analysis of DDX4 was performed by loading 25 µg of indicated whole cell lysates and 7 µl of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. DDX4 was detected at ~75-79 kDa using a DDX4 mouse monoclonal antibody (Product # MA5-15565, upper) at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform, followed by a Goat anti-mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 31430) at a dilution of 1:20,000 for at least 30 minutes at room temperature. GAPDH was detected using GAPDH rabbit polyclonal antibody (Product # PA1-987, lower), followed by a Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036) at a dilution of 1:2000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).

Product Details

MA5-15565

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:2,000

View 2 publications 2 publications

Immunohistochemistry (IHC)

1:200-1:1,000

Immunocytochemistry (ICC/IF)

1:100-1:200

View 1 publication 1 publication

Flow Cytometry (Flow)

1:200-1:400

Product Specifications

Species Reactivity

Human

Published species

Human, Protozoa

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

2F9H5

Immunogen

Purified recombinant fragment of human DDX4 expressed in E. Coli.

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A/G

Storage buffer

PBS

Contains

0.05% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_11001446

Product Specific Information

MA5-15565 targets DDX4 in FACS, IF, IHC, and WB applications and shows reactivity with Human samples.

The MA5-15565 immunogen is purified recombinant fragment of human DDX4 expressed in E. Coli.

MA5-15565 detects DDX4 which has a predicted molecular weight of approximately 76kDa.

Target Information

DEAD box proteins, characterized by the conserved motif Asp-Glu-Ala-Asp, are putative RNA helicases. They are implicated in a number of cellular processes involving alteration of RNA secondary structure such as translation initiation, nuclear and mitochondrial splicing, and ribosome and spliceosome assembly. Based on their distribution patterns, some members of this family are believed to be involved in embryogenesis, spermatogenesis, and cellular growth and division. This gene encodes a DEAD box protein, which is a homolog of VASA proteins in Drosophila and several other species. The gene is specifically expressed in the germ cell lineage in both sexes and functions in germ cell development. Multiple transcript variants encoding different isoforms have been found for this gene.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

"Mitotic Slippage" and Extranuclear DNA in Cancer Chemoresistance: A Focus on Telomeres.

International journal of molecular sciences

Salmina K,Bojko A,Inashkina I,Staniak K,Dudkowska M,Podlesniy P,Rumnieks F,Vainshelbaum NM,Pjanova D,Sikora E,Erenpreisa J

MA5-15565 was used in Western Blotting to explore mitotic slippage in the MDA-MB-231 cell line treated with doxorubicin.

Thu Apr 16 00:00:00 EDT 2020

"Mitotic Slippage" and Extranuclear DNA in Cancer Chemoresistance: A Focus on Telomeres.

International journal of molecular sciences

Salmina K,Bojko A,Inashkina I,Staniak K,Dudkowska M,Podlesniy P,Rumnieks F,Vainshelbaum NM,Pjanova D,Sikora E,Erenpreisa J

MA5-15565 was used in Western Blotting to explore mitotic slippage in the MDA-MB-231 cell line treated with doxorubicin.

Thu Apr 16 00:00:00 EDT 2020

Nuclear Factor Y (NF-Y) Modulates Encystation in Entamoeba via Stage-Specific Expression of the NF-YB and NF-YC Subunits.

mBio

Manna D,Singh U

MA5-15565 was used in Western Blot to demonstrate that NF-Y subunits in the protist Entamoeba, while significantly divergent from those of higher eukaryotes, have well-conserved domains important for subunit interactions and DNA binding and that NF-YB and NF-YC are developmentally expressed during encystation.

Tue Jun 18 00:00:00 EDT 2019

Bioinformatics

Protein Aliases: DEAD (Asp-Glu-Ala-Asp) box polypeptide 4; DEAD box protein 4; DEAD/H (Asp-Glu-Ala-Asp/His) box polypeptide 4; MGC111074; Probable ATP-dependent RNA helicase DDX4; Vasa homolog

Gene Aliases: DDX4; VASA

UniProt ID: (Human) Q9NQI0

Entrez Gene ID: (Human) 54514

Function(s)

male meiosis multicellular organism development spermatogenesis RNA secondary structure unwinding cell differentiation sperm motility gene silencing by RNA piRNA metabolic process DNA methylation involved in gamete generation

Process(es)

nucleic acid binding ATP-dependent RNA helicase activity ATP binding