Anti-FANCA/FAA antibody [EPR16519]

• IP

Supplier Data

Immunoprecipitation - Anti-FANCA/FAA antibody [EPR16519] (AB201457)

FANCA/FAA was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab201457 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201457 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (Input). Lane 2 : ab201457 IP in HeLa whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab201457 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-FANCA/FAA antibody [EPR16519] (ab201457)

Predicted band size: 163 kDa

false

• WB

Supplier Data

Western blot - Anti-FANCA/FAA antibody [EPR16519] (AB201457)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FANCA/FAA antibody [EPR16519] (ab201457) at 1/1000 dilution

All lanes:

Human colon 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: 163 kDa

Observed band size: 163 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-FANCA/FAA antibody [EPR16519] (AB201457)

Lanes 1, 5 and 9 : Wild-type HAP1 cell lysate (20 μg)
Lanes 2, 6 and 10 : FANCA/FAA beta knockout HAP1 cell lysate (20 μg)
Lanes 3, 7 and 11 : HeLa cell lysate (20 μg)
Lanes 4, 8 and 12 : HEK293 cell lysate (20 μg)
Lanes 1, 2, 3 and 4 : Green signal from target – ab201457 observed at 163 kDa
Lanes 5, 6, 7 and 8 : Red signal from loading control – ab8245 observed at 37 kDa
Lanes 9, 10, 11 and 12 : Merged (red and green) signal

ab201457 was shown to specifically react with FANCA/FAA beta when FANCA/FAA beta knockout samples were used. Wild-type and FANCA/FAA beta knockout samples were subjected to SDS-PAGE. ab201457 and ab8245 (loading control to GAPDH) were diluted 1/10000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-FANCA/FAA antibody [EPR16519] (ab201457)

Predicted band size: 163 kDa

false

• WB

Lab

Western blot - Anti-FANCA/FAA antibody [EPR16519] (AB201457)

Western blot : Anti-FANCA antibody [EPR16519] (ab201457) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab201457 was shown to bind specifically to FANCA. A band was observed at 163 kDa in wild-type A549 cell lysates with no signal observed at this size in FANCA knockout cell line. To generate this image, wild-type and FANCA 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.

Lanes 1 - 4:

Western blot - Anti-FANCA/FAA antibody [EPR16519] (ab201457) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-FANCA/FAA antibody [EPR16519] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/fanca-faa-antibody-epr16519-bsa-and-azide-free-ab251344'>ab251344</a>)

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

FANCA knockout A549 cell lysate at 20 µg

Lane 3:

Wild-type HAP1 cell lysate at 20 µg

Lane 4:

FANCA knockout HAP1 cell lysate at 20 µg

Observed band size: 163 kDa

false

• WB

Supplier Data

Western blot - Anti-FANCA/FAA antibody [EPR16519] (AB201457)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FANCA/FAA antibody [EPR16519] (ab201457) at 1/10000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate at 20 µg

Lane 3:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

Lane 4:

A431 (Human epidermoid carcinoma) whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 163 kDa

Observed band size: 163 kDa

false

Exposure time: 1min