Anti-USP30 antibody [EPR27024-81] - BSA and Azide free

• WB

Lab

Western blot - Anti-USP30 antibody [EPR27024-81] - BSA and Azide free (AB314750)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab314749).

Western blot : Rabbit Monoclonal [EPR27024-81] to USP30 ab314749 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 59 kDa in Wild-type RAW 264.7 cell lysates with no signal observed at this size in USP30 knockout RAW 264.7 cell line. To generate this image, 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-USP30 antibody [EPR27024-81] (<a href='/ko/products/primary-antibodies/usp30-antibody-epr27024-81-ab314749'>ab314749</a>) at 1/1000 dilution

Lane 1:

Wild-type RAW 264.7 cell lysates at 20 µg

Lane 2:

USP30 knockout RAW 264.7 cell lysates at 20 µg

Lane 3:

Wild-type A549 cell lysates at 20 µg

Lane 4:

USP30 knockout A549 <a href='/ko/products/cell-lines/human-usp30-knockout-a549-cell-line-ab324260'>ab324260</a> cell lysates at 20 µg

Secondary

All lanes:

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

Predicted band size: 59 kDa

Observed band size: 59 kDa

false

• WB

Supplier Data

Western blot - Anti-USP30 antibody [EPR27024-81] - BSA and Azide free (AB314750)

This data was developed using ab314749, the same antibody clone in a different buffer formulation.

ab314749 was shown to react with USP30 in wild-type MCF-7 cells in Western blot with loss of signal observed in USP30 knockout cell line ab324541. Wild-type MCF-7 and USP30 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab314749 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-USP30 antibody [EPR27024-81] (<a href='/ko/products/primary-antibodies/usp30-antibody-epr27024-81-ab314749'>ab314749</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF-7 lysate at 30 µg

Lane 2:

USP30 knock-out MCF-7 lysate at 30 µg

Lane 2:

Western blot - Human USP30 knockout MCF7 cell line (<a href='/ko/products/cell-lines/human-usp30-knockout-mcf7-cell-line-ab324541'>ab324541</a>)

Observed band size: 60 kDa

false

• WB

Supplier Data

Western blot - Anti-USP30 antibody [EPR27024-81] - BSA and Azide free (AB314750)

This data was developed using ab314749, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. This blot was developed using a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-USP30 antibody [EPR27024-81] (<a href='/ko/products/primary-antibodies/usp30-antibody-epr27024-81-ab314749'>ab314749</a>) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 58 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-USP30 antibody [EPR27024-81] - BSA and Azide free (AB314750)

This data was developed using ab314749, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-USP30 antibody [EPR27024-81] (<a href='/ko/products/primary-antibodies/usp30-antibody-epr27024-81-ab314749'>ab314749</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically target USP30 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 58 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-USP30 antibody [EPR27024-81] - BSA and Azide free (AB314750)

This data was developed using ab314749, the same antibody clone in a different buffer formulation. Western blot : Rabbit Monoclonal [EPR27024-81] to USP30 ab314749 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 59 kDa in Wild-type A549 ab288558 cell lysates with no signal observed at this size in USP30 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-USP30 antibody [EPR27024-81] (<a href='/ko/products/primary-antibodies/usp30-antibody-epr27024-81-ab314749'>ab314749</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human USP30 Knockout A549 cell line (<a href='/ko/products/cell-lines/human-usp30-knockout-a549-cell-line-ab324260'>ab324260</a>) at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 59 kDa

Observed band size: 59 kDa

false

• WB

Supplier Data

Western blot - Anti-USP30 antibody [EPR27024-81] - BSA and Azide free (AB314750)

This data was developed using ab314749, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. This blot was developed using a high sensitivity ECL substrate. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-USP30 antibody [EPR27024-81] (<a href='/ko/products/primary-antibodies/usp30-antibody-epr27024-81-ab314749'>ab314749</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 20 µg

Lane 3:

Rat brain tissue lysate at 20 µg

Lane 4:

Rat liver tissue lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/2000 dilution

Lanes 3 - 4:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/100000 dilution

Observed band size: 58 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-USP30 antibody [EPR27024-81] - BSA and Azide free (AB314750)

This data was developed using ab314749, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. This blot was developed using a high sensitivity ECL substrate. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-USP30 antibody [EPR27024-81] (<a href='/ko/products/primary-antibodies/usp30-antibody-epr27024-81-ab314749'>ab314749</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 5:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

T47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

Lane 7:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 58 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-USP30 antibody [EPR27024-81] - BSA and Azide free (AB314750)

This data was developed using ab314749, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. This blot was developed using a high sensitivity ECL substrate. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-USP30 antibody [EPR27024-81] (<a href='/ko/products/primary-antibodies/usp30-antibody-epr27024-81-ab314749'>ab314749</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 58 kDa

true

Exposure time: 180s