重组Anti-p73抗体[EP436Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p73 antibody [EP436Y] (AB40658)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver carcinoma tissue labelling p73 with purified ab40658 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-p73 antibody [EP436Y] (AB40658)

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling p73 with purified ab40658 at a dilution of 1/300. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1 : primary antibody (1/300) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2 : ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-p73 antibody [EP436Y] (AB40658)

Intracellular Flow Cytometry analysis of 293 (human embryonic kidney) cells labeling p73 with purified ab40658 at 1/120 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p73 antibody [EP436Y] (AB40658)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling p73 with purified ab40658 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p73 antibody [EP436Y] (AB40658)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human urinary bladder carcinoma tissue labelling p73 with unpurified ab40658 at a dilution of 1/100.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p73 antibody [EP436Y] (AB40658)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue labelling p73 with purified ab40658 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• WB

Lab

Western blot - Anti-p73 antibody [EP436Y] (AB40658)

Western blot : Anti-p73 antibody [EP436Y] ab40658 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 75 kDa in Wild-type HAP1 cell lysates with no signal observed at this size in TP73 knockout HAP1 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-p73 antibody [EP436Y] (ab40658) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysates at 20 µg

Lane 2:

TP73 knockout HAP1 cell lysates at 20 µg

Lane 3:

HEK-293 cell lysates at 20 µg

Lane 4:

HeLa cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 69 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-p73 antibody [EP436Y] (AB40658)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-p73 antibody [EP436Y] (ab40658) at 1/5000 dilution

Lane 1:

HeLa whole cell lysate at 10 µg

Lane 2:

HEK293 whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 69 kDa

Observed band size: 60 kDa,75 kDa

false

• WB

Lab

Western blot - Anti-p73 antibody [EP436Y] (AB40658)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-p73 antibody [EP436Y] (ab40658) at 1/1000 dilution

All lanes:

NIH/3T3 whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 69 kDa

Observed band size: 60 kDa

false

• WB

Unknown

Western blot - Anti-p73 antibody [EP436Y] (AB40658)

All lanes:

Western blot - Anti-p73 antibody [EP436Y] (ab40658) at 1/2000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Lane 3:

NIH/3T3 cell lysate at 10 µg

Predicted band size: 69 kDa

Observed band size: 63 kDa

false