Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal (AB196649)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling NR0B1 / Dax1 with ab196649 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution and counter-stained with Hematoxylin. (inset : negative control).

Note : Nuclear staining on human testis tissue was observed.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal (AB196649)

Intracellular Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling NR0B1 / Dax1 with purified ab196649 at 1/150 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal (AB196649)

Immunofluorescent analysis of SH-SY5Y cells (4% Paraformaldehyde-fixed 0.1% tritonX-100 permeabilized) labeling NR0B1 / Dax1 with ab196649 at 1/300 dilution followed by Goat anti rabbit IgG (AlexaFluor® 488) (ab150077) secondary at 1/400 dilution and counter-stained with DAPI (blue).

Note : Nuclear and cytoplasm staining on SH-SY5Y cell line was observed.

• WB

Lab

Western blot - Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal (AB196649)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of A673 whole cell lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab196649 (1/2000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/20,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique. ab196649 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected.

All lanes:

Western blot - Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal (ab196649) at 1/2000 dilution

All lanes:

A673 whole cell lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

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Exposure time: 7s

• WB

Supplier Data

Western blot - Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal (AB196649)

All lanes:

Western blot - Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal (ab196649) at 1/1000 dilution

All lanes:

A431 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 52 kDa

false

• WB

Supplier Data

Western blot - Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal (AB196649)

All lanes:

Western blot - Anti-NR0B1 / Dax1 antibody [EP13786] - N-terminal (ab196649) at 1/1000 dilution

All lanes:

Human testis lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 52 kDa

false