Anti-DDX4 / MVH antibody [mAbcam27591]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX4 / MVH antibody [mAbcam27591] (AB27591)

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 ab27591, 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

• IHC-P

AbReview33758****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX4 / MVH antibody [mAbcam27591] (AB27591)

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).

This image is courtesy of an abreview from Zachary Yu-Ching Lin.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX4 / MVH antibody [mAbcam27591] (AB27591)

Immunohistochemical analysis of formalin fixed paraffin embedded human testis labelling DDX4/MVH with ab27591 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab27591 anti-DDX4/MVH antibody [mAbcam27591] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DDX4 / MVH antibody [mAbcam27591] (AB27591)

ab27591 staining DDX4/MVH in mES cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab27591 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DDX4 / MVH antibody [mAbcam27591] (AB27591)

ab27591 staining DDX4/MVH in mES cells. The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab27591 at 10μg/ml concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI. Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

• WB

Project4670****

Western blot - Anti-DDX4 / MVH antibody [mAbcam27591] (AB27591)

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.

All lanes:

Western blot - Anti-DDX4 / MVH antibody [mAbcam27591] (ab27591) at 5 µg/mL

Lane 1:

Mouse Testis Tissue Lysate at 20 µg

Lane 2:

Mouse EG (TMAS Embryonic Germ Cells) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-preadsorbed-ab97040'>ab97040</a>) at 1/5000 dilution

Predicted band size: 79 kDa

Observed band size: 70 kDa,80 kDa

true

Exposure time: 1min

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-DDX4 / MVH antibody [mAbcam27591] (AB27591)

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. ab150110 at a dilution of 1/500 was used as the secondary antibody.