Anti-PSMB10/MECL1 antibody [EPR14902] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PSMB10/MECL1 antibody [EPR14902] - BSA and Azide free (AB240272)

Immunohistochemical analysis of paraffin-embedded Human Kidney adenocarcinoma tissue labeling PSMB10/MECL1 with ab183506 at 1/100 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183506).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PSMB10/MECL1 antibody [EPR14902] - BSA and Azide free (AB240272)

Intracellular flow cytometric analysis of Daudi cells (2% paraformaldehyde-fixed) labeling PSMB10/MECL1 with ab183506 at 1/90 dilution (red) or a rabbit IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183506).

• WB

Lab

Western blot - Anti-PSMB10/MECL1 antibody [EPR14902] - BSA and Azide free (AB240272)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183506).

Western blot : Anti-PSMB10/MECL1 antibody [EPR14902] ab183506 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 25,29 kDa in Wild-type HeLa cell lysates with no signal observed at this size in PSMB10 knockout HeLa 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-PSMB10/MECL1 antibody [EPR14902] (<a href='/ko/products/primary-antibodies/psmb10-mecl1-antibody-epr14902-ab183506'>ab183506</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysates at 40 µg

Lane 2:

Western blot - Human PSMB10 (MECL1) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-psmb10-mecl1-knockout-hela-cell-line-ab264923'>ab264923</a>) at 40 µg

Lane 3:

Daudi cell lysates at 20 µg

Lane 4:

Raji cell lysates at 20 µg

Secondary

All lanes:

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

Predicted band size: 29 kDa

Observed band size: 29 kDa,25 kDa

false

• WB

Lab

Western blot - Anti-PSMB10/MECL1 antibody [EPR14902] - BSA and Azide free (AB240272)

This data was developed using the same antibody clone in a different buffer formulation (ab183506).

Lanes 1-4 : Merged signal (red and green). Green - ab183506 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa.

ab183506 Anti-PSMB10/MECL1 antibody [EPR14902] was shown to specifically react with PSMB10/MECL1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264923 (knockout cell lysate ab258145) was used. Wild-type and PSMB10/MECL1 knockout samples were subjected to SDS-PAGE. ab183506 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PSMB10/MECL1 antibody [EPR14902] (<a href='/ko/products/primary-antibodies/psmb10-mecl1-antibody-epr14902-ab183506'>ab183506</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PSMB10 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PSMB10 (MECL1) knockout HeLa cell line (<a href='/ko/products/cell-lines/human-psmb10-mecl1-knockout-hela-cell-line-ab264923'>ab264923</a>)

Lane 3:

Daudi cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 29 kDa

Observed band size: 29 kDa

false