Immunohistochemistry (Frozen sections) - Anti-DDX4 / MVH antibody - Primordial Germ Cell Marker (ab13840)

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.

Petra Hajkova, Gurdon Institute, University of Cambridge

Western blot - Anti-DDX4 / MVH antibody - Primordial Germ Cell Marker (ab13840)

All lanes : Anti-DDX4 / MVH antibody - Primordial Germ Cell Marker (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) - DDX4 / MVH antibody - Primordial Germ Cell Marker (ab13840)

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.

This image is courtesy of an anonymous Abreview

Immunocytochemistry/ Immunofluorescence - DDX4 / MVH antibody - Primordial Germ Cell Marker (ab13840)

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.

This image is courtesy of an Abreview submitted by Mai Sarraj

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - DDX4 / MVH antibody - Primordial Germ Cell Marker (ab13840)

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

This image is courtesy of an Abreview submitted by Mr kyle caires

Western blot - DDX4 / MVH antibody - Primordial Germ Cell Marker (ab13840)

All lanes : Anti-DDX4 / MVH antibody - Primordial Germ Cell Marker (ab13840) at 1 µg/ml

Lane 1 : Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate
Lane 2 : Testis (Human) Tissue Lysate - adult normal tissue (ab30257)

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 76 kDa
Observed band size : 76 kDa
Additional bands at : 39 kDa. We are unsure as to the identity of these extra bands.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - DDX4 / MVH antibody - Primordial Germ Cell Marker (ab13840)

IHC image of DDX4/MVH staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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.