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.

Secondary
All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 76 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?
Additional bands at: 70 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 1 minute


Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

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).

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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.

IHC image of DDX4/MVH antibody staining in a section of formalin-fixed paraffin-embedded normal human testis* performed on a Leica BOND^TM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab27561, 1ug/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.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre