Anti-HNRNPA0 antibody [EP16085] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HNRNPA0 antibody [EP16085] - BSA and Azide free (AB236121)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling HNRNPA0 with ab197023 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
-ve control 1 : ab197023 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab197023).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNRNPA0 antibody [EP16085] - BSA and Azide free (AB236121)

Immunohistochemical analysis of paraffin-embedded Human cerebral cortex labeling HNRNPA0 with ab197023 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human cerebral cortex tissue is observed. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab197023).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-HNRNPA0 antibody [EP16085] - BSA and Azide free (AB236121)

Intracellular Flow Cytometry analysis of HeLa cells labelling HNRNPA0 (red) with purified ab197023 at dilution of 1/150. The secondary antibody used was Alexa Fluor^® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab197023).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNRNPA0 antibody [EP16085] - BSA and Azide free (AB236121)

Immunohistochemical analysis of paraffin-embedded Mouse spleen labeling HNRNPA0 with ab197023 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Mouse spleen tissue is observed. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab197023).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-HNRNPA0 antibody [EP16085] - BSA and Azide free (AB236121)

This data was developed using the same antibody clone in a different buffer formulation (ab197023).

Lanes 1-3 : Merged signal (red and green). Green - ab197023 observed at 32,34 kDa. Red - loading control ab8245 observed at 36 kDa.

ab197023 Anti-HNRNPA0 antibody [EP16085] was shown to specifically react with HNRNPA0 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266314 (knockout cell lysate ab257989) was used. Wild-type and HNRNPA0 knockout samples were subjected to SDS-PAGE. ab197023 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HNRNPA0 antibody [EP16085] (<a href='/en-us/products/primary-antibodies/hnrnpa0-antibody-ep16085-ab197023'>ab197023</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

HNRNPA0 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human HNRNPA0 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-hnrnpa0-knockout-hek-293t-cell-line-ab266314'>ab266314</a>)

Lane 3:

A549 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 31 kDa

Observed band size: 32 kDa,34 kDa

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