Anti-RAB10 antibody [EPR13242] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RAB10 antibody [EPR13242] - BSA and Azide free (AB224886)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RAB10 with Purified ab181367 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181367).

• WB

Lab

Western blot - Anti-RAB10 antibody [EPR13242] - BSA and Azide free (AB224886)

Lanes 1 - 4 : Merged signal (red and green). Green - ab181367 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab181367 was shown to specifically react with RAB10 in wild-type A549 cells as signal was lost in RAB10 knockout cells. Wild-type and RAB10 knockout samples were subjected to SDS-PAGE. ab181367 and ab8245(Mouse anti GAPDH 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181367).

All lanes:

Western blot - Anti-RAB10 antibody [EPR13242] (<a href='/en-us/products/primary-antibodies/rab10-antibody-epr13242-ab181367'>ab181367</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

RAB10 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human RAB10 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-rab10-knockout-a549-cell-line-ab261868'>ab261868</a>)

Lane 3:

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

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 25 kDa

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• WB

Lab

Western blot - Anti-RAB10 antibody [EPR13242] - BSA and Azide free (AB224886)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181367).

All lanes:

Western blot - Anti-RAB10 antibody [EPR13242] (<a href='/en-us/products/primary-antibodies/rab10-antibody-epr13242-ab181367'>ab181367</a>) at 1/1000 dilution

Lane 1:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 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: 22 kDa

Observed band size: 22 kDa

false

• WB

Lab

Western blot - Anti-RAB10 antibody [EPR13242] - BSA and Azide free (AB224886)

This data was developed using ab181367, the same antibody clone in a different buffer formulation.

ab181367 was shown to react with RAB10 in wild-type HAP1 cells in Western blot with loss of signal observed in a RAB10 knockout cell line. Wild-type HAP1 and RAB10 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab181367 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were 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-RAB10 antibody [EPR13242] (<a href='/en-us/products/primary-antibodies/rab10-antibody-epr13242-ab181367'>ab181367</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 lysate at 40 µg

Lane 2:

RAB10 knock-out HAP1 lysate at 40 µg

false

• WB

Lab

Western blot - Anti-RAB10 antibody [EPR13242] - BSA and Azide free (AB224886)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181367).

All lanes:

Western blot - Anti-RAB10 antibody [EPR13242] (<a href='/en-us/products/primary-antibodies/rab10-antibody-epr13242-ab181367'>ab181367</a>) at 1/2000 dilution

Lane 1:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

Lane 2:

C6 (Rat glial tumor glial cell) whole cell lysates at 15 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: 22 kDa

Observed band size: 22 kDa

false