Anti-RAB13 antibody [EPR14110(2)] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RAB13 antibody [EPR14110(2)] - BSA and Azide free (AB251431)

This data was developed using ab205528, the same antibody clone in a different buffer formulation.

ab205528 was shown to react with RAB13 in wild-type U-87 MG cells in immunocytochemistry with loss of signal observed in a RAB13 siRNA knockdown cell line. Wild-type and siRNA knockdown cells were mixed and pelleted at a 1 : 1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab205528 at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor^® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (siRNA knockdown) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and siRNA knockdown cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

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.

• WB

Supplier Data

Western blot - Anti-RAB13 antibody [EPR14110(2)] - BSA and Azide free (AB251431)

This data was developed using ab205528, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RAB13 antibody [EPR14110(2)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr141102-ab205528'>ab205528</a>) at 1/2000 dilution

Lane 1:

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

Lane 2:

Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) 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 1/100000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

false

Exposure time: 1min

• WB

Lab

Western blot - Anti-RAB13 antibody [EPR14110(2)] - BSA and Azide free (AB251431)

This data was developed using ab205528, the same antibody clone in a different buffer formulation.

Lanes 1 - 4 : Merged signal (red and green). Green - ab205528 observed at 23 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab205528 was shown to specifically react with RAB13 in wild-type HAP1 cells as signal was lost in RAB13 knockout cells. Wild-type and RAB13 knockout samples were subjected to SDS-PAGE. ab205528 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 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.

All lanes:

Western blot - Anti-RAB13 antibody [EPR14110(2)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr141102-ab205528'>ab205528</a>) at 1/2000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

RAB13 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

HEK293 whole cell lysate at 20 µg

Predicted band size: 23 kDa

false

• WB

Supplier Data

Western blot - Anti-RAB13 antibody [EPR14110(2)] - BSA and Azide free (AB251431)

This data was developed using ab205528, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RAB13 antibody [EPR14110(2)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr141102-ab205528'>ab205528</a>) at 1/2000 dilution

All lanes:

Human fetal lung lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-RAB13 antibody [EPR14110(2)] - BSA and Azide free (AB251431)

This data was developed using ab205528, the same antibody clone in a different buffer formulation.

ab205528 was shown to react with RAB13 in wild-type U-87 MG cells in Western blot with loss of signal observed in a RAB13 siRNA knockdown cell line. Cell lysates from wild-type U-87 MG transfected with either scrambled siRNA or RAB13 siRNA were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hour before incubation with ab205528 overnight at 4 °C at a 1/200 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-RAB13 antibody [EPR14110(2)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr141102-ab205528'>ab205528</a>) at 1/200 dilution

Lane 1:

Wild-type U-87 MG transfected with scrambled siRNA control lysate at 20 µg

Lane 2:

U-87 MG transfected with siRNA specifically targeting RAB13 cell lysate at 20 µg

false

• WB

Supplier Data

Western blot - Anti-RAB13 antibody [EPR14110(2)] - BSA and Azide free (AB251431)

This data was developed using ab05528, the same antibody clone in a different buffer formulation.

ab205528 was shown to react with RAB13 in wild-type HEK-293T cells in Western blot with loss of signal observed in RAB13 knockout cell line ab266843. Wild-type HEK-293T and RAB13 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hour before incubation with ab205528 overnight at 4 °C at a 1/200 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-RAB13 antibody [EPR14110(2)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr141102-ab205528'>ab205528</a>) at 1/200 dilution

Lane 1:

Wild-type HEK-293T lysate at 30 µg

Lane 2:

RAB13 knock-out HEK-293T lysate at 30 µg

false

• WB

Lab

Western blot - Anti-RAB13 antibody [EPR14110(2)] - BSA and Azide free (AB251431)

This data was developed using ab205528, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab205528 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.

ab205528 Anti-RAB13 antibody [EPR14110(2)] was shown to specifically react with RAB13 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266843 (knockout cell lysate ab258159) was used. Wild-type and RAB13 knockout samples were subjected to SDS-PAGE. ab205528 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-RAB13 antibody [EPR14110(2)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr141102-ab205528'>ab205528</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

RAB13 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human RAB13 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-rab13-knockout-hek-293t-cell-line-ab266843'>ab266843</a>)

Lane 3:

HAP1 whole cell lyate at 20 µg

Lane 4:

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

Secondary

All lanes:

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

Predicted band size: 23 kDa

Observed band size: 24 kDa

false