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.
![All lanes : Anti-DDX4 / MVH antibody [mAbcam27591] (ab27591) at 5 µg/ml (Blocked with 3% Milk)Lane 1 : Mouse Testis Tissue Lysate Lane 2 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate Lysates/proteins at 20 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/7233_DDX4-MVH-Primary-antibodies-ab27591-12.jpg)
All lanes : Anti-DDX4 / MVH antibody [mAbcam27591] (ab27591) at 5 µg/ml (Blocked with 3% Milk)Lane 1 : Mouse Testis Tissue Lysate Lane 2 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate Lysates/proteins at 20 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
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.See Abreview
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.
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).See Abreview