This antibody gave a positive signal in Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate and Human Testis Tissue Lysate
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The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 76 kDa (predicted molecular weight: 76 kDa).Can be blocked with DDX4 / MVH peptide (ab13841).
Use at an assay dependent concentration. PubMed: 23598447
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
1/200. PubMed: 19440488
Use a concentration of 1 µg/ml.
Probable ATP-dependent RNA helicase required during spermatogenesis (PubMed:10920202, PubMed:21034600). Required to repress transposable elements and preventing their mobilization, which is essential for the germline integrity. Acts via the piRNA metabolic process, which mediates the repression of transposable elements during meiosis by forming complexes composed of piRNAs and Piwi proteins and governs the methylation and subsequent repression of transposons. Involved in the secondary piRNAs metabolic process, the production of piRNAs in fetal male germ cells through a ping-pong amplification cycle.
Expressed only in ovary and testis. Expressed in migratory primordial germ cells in the region of the gonadal ridge in both sexes.
Belongs to the DEAD box helicase family. DDX4/VASA subfamily. Contains 1 helicase ATP-binding domain. Contains 1 helicase C-terminal domain.
Cytoplasm. Cytoplasm, perinuclear region. Component of the meiotic nuage, also named P granule, a germ-cell-specific organelle required to repress transposon activity during meiosis.
Immunohistochemistry (Frozen sections) - Anti-DDX4 / MVH antibody (ab13840)This image is courtesy of Petra Hajkova, Gurdon Institute, University of Cambridge
Mouse embryonic gonad cells were fixed with 4% PFA, permeabilised and blocked with PBS, 1% BSA, 0.1% TX-100 and incubated with the DDX4 antibody, ab13840 (1 µg/ml) overnight at 4 degrees. ab13840 staining is shown in red and Oct4 staining is shown in green.
The image shows that the anti-DDX4 antibody, ab13840, stains the cytoplasm of mouse primordial germ cells when used in IHC-Fr.
Immunocytochemistry/ Immunofluorescence - Anti-DDX4 / MVH antibody (ab13840)This image is courtesy of an Abreview submitted by Mai Sarraj
ab13840 at 1/1000 staining germ cells of the embryonic mouse testis by ICC/IF. Cells were PFA fixed and permeabilized in 1% Triton X in PBS prior to blocking in 5% serum for 2 hours at 20°C. An Alexa Fluor® 488 conjugated goat anti-rabbit antibody was used as the secondary.
All lanes : Anti-DDX4 / MVH antibody (ab13840) at 1 µg/ml
Lane 1 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate Lane 2 : Human testis tissue lysate - total protein (ab30257)
Lysates/proteins at 20 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 76 kDa Observed band size: 76 kDa Additional bands at: 28 kDa, 47 kDa. We are unsure as to the identity of these extra bands.
Western blot - Anti-DDX4 / MVH antibody (ab13840)
All lanes : Anti-DDX4 / MVH antibody (ab13840) at 1 µg/ml
Lane 1 : Mouse Ovary Lysate at 10 µg Lane 2 : Mouse Embryonic Germ Cell Lysate at 20 µg Lane 3 : Mouse Ovary Lysate at 10 µg with DDX4 / MVH peptide (ab13841) at 1 µg/ml Lane 4 : Mouse Embryonic Germ Cell Lysate at 20 µg with DDX4 / MVH peptide (ab13841) at 1 µg/ml
Predicted band size: 76 kDa Observed band size: 76 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX4 / MVH antibody (ab13840)This image is courtesy of an anonymous Abreview
Ab13840 at 1/400 staining mouse fetal testis tissue sections by IHC-P. The tissue was blocked with both BSA and serum and fixed prior to incubation with the antibody for 3 hours at room temperature. A biotinylated secondary antibody was used (Vectastain Elite).
Please note Bouin's fluid was used as the fixative in this experiment.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX4 / MVH antibody (ab13840)This image is courtesy of an Abreview submitted by Mr Kyle Caires
ab13840 staining Postnatal Porcine Testis tissue sections by IHC-P. Bouin's Fixative was used to fix the sections prior to a heat mediated antigen retrieval step in sodium citrate buffer (pH 6). The sample was blocked in 10% serum for 1 hour at 22°C prior to incubation with the primary antibody, diluted 1/500, for 12 hours at 4°C. A biotinylated goat anti-rabbit antibody was used as the secondary
IHC image of DDX4/MVH staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab13840, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Magaraki A et al. Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development. Epigenetics Chromatin10:11 (2017).
Read more (PubMed: 28293300) »
Liu W et al. BCAS2 is involved in alternative mRNA splicing in spermatogonia and the transition to meiosis. Nat Commun8:14182 (2017).
WB, Flow Cyt, ICC/IF
Read more (PubMed: 28128212) »