Anti-PHAP1 antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PHAP1 antibody (AB5992)

Immunocytochemical analysis of Raji cells labeling PHAP1 with ab5992 at 2 μg/mL. Cells were fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PHAP1 antibody (AB5992)

Immunofluorescent analysis of 4% paraformaldehydefixed Raji Cells labeling PHAP I with ab5992 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

• WB

Lab

Western blot - Anti-PHAP1 antibody (AB5992)

False colour image of Western blot : Anti-PHAP1 antibody staining at 0.5 ug/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab5992 was shown to bind specifically to PHAP1. A band was observed at 33 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in ANP32A knockout cell line ab266148 (knockout cell lysate ab258303). To generate this image, wild-type and ANP32A knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L (IRDye^� 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^� 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Lanes 1 - 4:

Western blot - Anti-PHAP1 antibody (<a href='/en-us/products/primary-antibodies/phap1-antibody-ab5991'>ab5991</a>) at 0.5 µg/mL

Lanes 1 - 4:

Western blot - Anti-PHAP1 antibody (ab5992) at 0.5 µg/mL

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

ANP32A knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human ANP32A (PHAP1) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-anp32a-phap1-knockout-hek-293t-cell-line-ab266148'>ab266148</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

THP-1 cell lysate at 20 µg

Predicted band size: 28 kDa

Observed band size: 33 kDa

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• WB

Unknown

Western blot - Anti-PHAP1 antibody (AB5992)

Western blot analysis of PHAP I expression in human Raji cell lysate with anti-PHAP I at 2 μg/ml (lane A) and 4 μg/ml (lane B), respectively. Western blot analysis of PHAP I expression in human Raji cell lysate with anti-PHAP I at 2 µg/ml (lane A) and 4 µg/ml (lane B), respectively.

All lanes:

Western blot - Anti-PHAP1 antibody (ab5992)

Predicted band size: 28 kDa

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• WB

Supplier Data

Western blot - Anti-PHAP1 antibody (AB5992)

All lanes:

Western blot - Anti-PHAP1 antibody (ab5992) at 2 µg/mL

Lane 1:

Daudi cell lysates at 15 µg

Lane 2:

HeLa cell lysates at 15 µg

Lane 3:

HepG2 cell lysates at 15 µg

Lane 4:

K562 cell lysates at 15 µg

Secondary

All lanes:

Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

Predicted band size: 28 kDa

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