Anti-FAM38A/PIEZO1 antibody [EPR23826-103]

• WB

Lab

Western blot - Anti-FAM38A/PIEZO1 antibody [EPR23826-103] (AB259949)

Western blot : Anti-FAM38A/PIEZO1 antibody [EPR23826-103] ab259949 staining at 1/1000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 240-350 kDa in Wild-type A549 not boiled cell lysates with no signal observed at this size in PIEZO1 knockout A549 not boiled cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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-FAM38A/PIEZO1 antibody [EPR23826-103] (ab259949) at 1/1000 dilution

Lane 1:

Wild-type A549 not boiled at 20 µg

Lane 2:

PIEZO1 knockout A549 not boiled at 20 µg

Lane 3:

Wild-type HeLa not boiled at 20 µg

Lane 4:

PIEZO knockout HeLa not boiled at 20 µg

Lane 5:

A431 not boiled at 20 µg

Secondary

All lanes:

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

Predicted band size: 240-350 kDa

Observed band size: 240-350 kDa

false

• WB

Lab

Western blot - Anti-FAM38A/PIEZO1 antibody [EPR23826-103] (AB259949)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 29396846).

Exposure time : 74 seconds.

All lanes:

Western blot - Anti-FAM38A/PIEZO1 antibody [EPR23826-103] (ab259949) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 4.5 µg

Lane 2:

HeLa transfected with siRNA specifically targeting PIEZO1 whole cell lysate at 5 µg

Secondary

All lanes:

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

Predicted band size: 287 kDa

Observed band size: 300 kDa

false

• WB

Lab

Western blot - Anti-FAM38A/PIEZO1 antibody [EPR23826-103] (AB259949)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID : 29396846).

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-FAM38A/PIEZO1 antibody [EPR23826-103] (ab259949) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell), transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

A431 transfected with 25nM siRNA specifically targeting PIEZO1 whole cell lysate at 20 µg

Lane 3:

A431 transfected with 100nM siRNA specifically targeting PIEZO1 whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 287 kDa

Observed band size: 300 kDa

false

• WB

Lab

Western blot - Anti-FAM38A/PIEZO1 antibody [EPR23826-103] (AB259949)

Western blot : Anti-PIEZO1 antibody [EPR23826-103] (ab259949) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab259949 was shown to bind specifically to PIEZO1. A band was observed at 286 kDa in wild-type HeLa cell lysates with no signal observed at this size in PIEZO1 knockout cell line. To generate this image, wild-type and PIEZO1 knockout HeLa cell lysates were analysed. First, 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 IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-FAM38A/PIEZO1 antibody [EPR23826-103] (ab259949) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PIEZO1 knockout HeLa cell lysate at 20 µg

Lane 3:

A431 UNBOILED cell lysate at 20 µg

Lane 4:

THP-1 UNBOILED cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Predicted band size: 287 kDa

Observed band size: 286 kDa

false