Anti-Annexin V/ANXA5 antibody [EPR3979]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Annexin V/ANXA5 antibody [EPR3979] (AB108321)

Immunohistochemical analysis of Annexin V/ANXA5 in paraffin embedded Human gastric tissue using ab108321 at a 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-Annexin V/ANXA5 antibody [EPR3979] (AB108321)

All lanes:

Western blot - Anti-Annexin V/ANXA5 antibody [EPR3979] (ab108321) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

JAR cell lysate at 10 µg

Lane 3:

fetal kidney lysate at 10 µg

Predicted band size: 35 kDa

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• WB

Lab

Western blot - Anti-Annexin V/ANXA5 antibody [EPR3979] (AB108321)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : Annexin V/ANXA5 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)

Lanes 1 - 3 : Merged signal (red and green). Green - ab108321 observed at 35 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab108321 was shown to specifically react with Annexin V/ANXA5 in wild-type cells as signal was lost in Annexin V/ANXA5 knockout cells. Wild-type and Annexin V/ANXA5 knockout samples were subjected to SDS-PAGE. ab108321 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Annexin V/ANXA5 antibody [EPR3979] (ab108321)

Predicted band size: 35 kDa

false

• WB

Lab

Western blot - Anti-Annexin V/ANXA5 antibody [EPR3979] (AB108321)

Western blot : Anti-ANXA5 antibody [EPR3979] (ab108321) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab108321 was shown to bind specifically to ANXA5. A band was observed at 36 kDa in wild-type A549 cell lysates with no signal observed at this size in ANXA5 knockout cell line. To generate this image, wild-type and ANXA5 knockout A549 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Annexin V/ANXA5 antibody [EPR3979] (ab108321) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human ANXA5 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-anxa5-knockout-a549-cell-line-ab300840'>ab300840</a>)

Lane 2:

ANXA5 knockout A549 cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 36 kDa

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