The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 10 µg/ml.
Use at an assay dependent concentration.
Use a concentration of 0.05 - 0.1 µg/ml.
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 76 kDa).
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 [mAbcam27591] (ab27591)This image is courtesy of an Abreview by Bryan Niedenberger.
ab27591 staining DDX4 / MVH antibody in pig tissue sections (testis, postnatal day 42) by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% 0.1% Triton X-100 in PBS and blocked with 3% BSA for 30 minutes at 20°C. Samples were incubated with primary antibody (1/200 in PBS) for 1 hour at 20°C. Ab150110l at a dilution of 1/500 was used as the secondary antibody.
Exposure time : 1 minuteAbcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX4 / MVH antibody [mAbcam27591] (ab27591)This image is courtesy of an abreview from Zachary Yu-Ching Lin.
IHC-P image of DDX4 / MVH (ab27591) on adult marmoset testis. The sections were fixed with Formaldehyde and blocked in 5% milk for 30 minutes, before incubation with the DDX4 antibody (1/50 dilution) for 18 hours at 4°C. Cytoplasmic staining is seen with ab27591 (green), DAPI was used to stain nuclei (blue).
ICC/IF image of ab27591 stained mES cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab27591, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistry (Frozen sections) - Anti-DDX4 / MVH antibody [mAbcam27591] (ab27591)This image is courtesy of an anonymous abreview.
IHC-Fr image of DDX4 (ab27591) on mouse embryonic gonad cells. The sections were fixed with 4% PFA, permeabilized and blocked in BSA, before incubation with the DDX4 antibody (1/100 dilution) for 12 hours at 4 degrees. Cytoplasmic staining of the mouse primordial germ cells by ab27591 is shown in red and Oct4-GFP staining is shown in green.
IHC image of DDX4/MVH staining in human normal 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 ab27591, 0.1µg/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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Lopes F et al. Irinotecan metabolite SN38 results in germ cell loss in the testis but not in the ovary of prepubertal mice. Mol Hum Reprod22:745-755 (2016).
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