Anti-DDX4 / MVH antibody

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX4 / MVH antibody (AB13840)

Immunohistochemical analysis of formalin fixed paraffin embedded human testis labelling DDX4/MVH with ab13840 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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX4 / MVH antibody (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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DDX4 / MVH antibody (AB13840)

ab13840 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 ab13840 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed 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.

• IHC-Fr

Collaborator

Immunohistochemistry (Frozen sections) - Anti-DDX4 / MVH antibody (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.

This image is courtesy of Petra Hajkova, Gurdon Institute, University of Cambridge

• WB

Unknown

Western blot - Anti-DDX4 / MVH antibody (AB13840)

All lanes:

Western blot - Anti-DDX4 / MVH antibody (ab13840) at 1 µg/mL

Lane 1:

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

Lane 2:

Human testis tissue lysate - total protein (<a href='/en-us/products/unavailable/human-testis-tissue-lysate-total-protein-ab30257'>ab30257</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 79 kDa

Observed band size: 28 kDa,47 kDa,76 kDa

false

• WB

Lab

Western blot - Anti-DDX4 / MVH antibody (AB13840)

All lanes:

ab13840 at 1 µg/mL

Lane 1:

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

Lane 2:

Human testis tissue lysate - total protein at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 79 kDa

Observed band size: 47 kDa,52 kDa,79 kDa

true

• WB

Project

Western blot - Anti-DDX4 / MVH antibody (AB13840)

All lanes:

Western blot - Anti-DDX4 / MVH antibody (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 (<a href='/en-us/products/proteins-peptides/ddx4-mvh-peptide-ab13841'>ab13841</a>)

Lane 4:

Mouse Embryonic Germ Cell Lysate at 20 µg with DDX4 / MVH peptide (<a href='/en-us/products/proteins-peptides/ddx4-mvh-peptide-ab13841'>ab13841</a>)

Predicted band size: 79 kDa

Observed band size: 76 kDa

false

• WB

CiteAb

Western blot - Anti-DDX4 / MVH antibody (AB13840)

Western Blotting using Anti-DDX4 / MVH antibody, ab13840. Publication image from Gao, F. et al., 2017, Nat Commun, 28128212. Legend direct from paper.

Expression of BCAS2 in male mouse germ cells.(a) Immunofluorescence (IF) staining of BCAS2 in the paraffin sections of testes from E15.5 to P14 mice. The DNA was stained with Hoechst 33342. Scale bar, 50 µm. (b) Real-time PCR analysis of Bcas2 expression in the fraction of spermatogenic cells (FSPCs) and the fraction of somatic cells (FSCs) enriched from P9 testes. Gapdh was used as the internal control for normalization (n=4). Error bars represent s.e.m. (c) Western blotting analysis of BCAS2 expression in the fraction of spermatogenic cells (FSPCs) and the fraction of somatic cells (FSCs) enriched from P9 testes. Germ cell markers (DAZL and MVH) were used as the indicator of the enrichment efficiency andα-tubulin was used as the loading control. (d) Paraffin sections of P8 testes were co-stained with rabbit anti-BCAS2 and mouse anti-PLZF antibodies. The DNA was stained with Hoechst 33342. Scale bar, 20 µm.

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