Anti-RAB13 antibody [EPR14109(B)] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RAB13 antibody [EPR14109(B)] - BSA and Azide free (AB250269)

This image was produced using ab180936, the same clone but in a different formulation.

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell) (+ve cell line), and HL-60 (human acute promyelocytic leukemia promyeloblast) (Low expressing cell line) labelling RAB13 with ab180936 at 1/100 (10.75 μg/ml) followed by ab150077 Goat Anti-Rabbit (Alexa Fluor^® 488) as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing mainly cytoplasmic staining in U-87 MG cell line (shown in green). ab7291 Anti-alpha Tubulin antibody [DM1A] - Loading Control was used as a counterstain at 1/200 with ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed as secondary (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). Low expression : HL-60. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RAB13 antibody [EPR14109(B)] - BSA and Azide free (AB250269)

This image was produced using ab180936, the same clone but in a different formulation.

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized Wildtype HEK-293T (human embryonic kidney epithelial cell), and RAB13 KO HEK293T (RAB13 knockout HEK293T) cells labelling RAB13 with ab180936 at 1/100 (10.75 μg/ml) followed by ab150077 Goat Anti-Rabbit (Alexa Fluor^® 488) as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing cytoplasmic and membranous staining in wildtype HEK-293T (shown in green), showing no staining in RAB13 knockout HEK-293T cells. ab7291 Anti-alpha Tubulin antibody [DM1A] - Loading Control was used as a counterstain at 1/200 with ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed as secondary (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RAB13 antibody [EPR14109(B)] - BSA and Azide free (AB250269)

This data was developed using ab180936, the same antibody clone in a different buffer formulation.

ab180936 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 ab180936 at 1/600 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 ug/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.

• IP

Unknown

Immunoprecipitation - Anti-RAB13 antibody [EPR14109(B)] - BSA and Azide free (AB250269)

ab250269 used at 1 : 30 dilution (2μg) immunoprecipitating RAB13 in Hela whole cell lysates.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : HeLa whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab190685 in HeLa whole cell lysate

For western blotting, ab227648 and VeriBlot for IP Detection Reagent (HRP)(ab131366) at 1/1000 dilution were used.

All lanes:

Immunoprecipitation - Anti-RAB13 antibody [EPR14109(B)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr14109b-ab180936'>ab180936</a>)

Predicted band size: 23 kDa

false

• WB

Lab

Western blot - Anti-RAB13 antibody [EPR14109(B)] - BSA and Azide free (AB250269)

This data was developed using ab180936, the same antibody clone in a different buffer formulation.

Lanes 1 - 4 : Merged signal (red and green). Green - ab180936 observed at 23 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab180936 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. ab180936 and ab9484 (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.

All lanes:

Western blot - Anti-RAB13 antibody [EPR14109(B)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr14109b-ab180936'>ab180936</a>) at 1/1000 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 [EPR14109(B)] - BSA and Azide free (AB250269)

This data was developed using ab180936, the same antibody clone in a different buffer formulation.

ab180936 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 ab180936 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 [EPR14109(B)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr14109b-ab180936'>ab180936</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

Supplier Data

Western blot - Anti-RAB13 antibody [EPR14109(B)] - BSA and Azide free (AB250269)

This data was developed using ab180936, the same antibody clone in a different buffer formulation.

ab180936 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 ab180936 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 [EPR14109(B)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr14109b-ab180936'>ab180936</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 lyse at 20 µg

false

• WB

Lab

Western blot - Anti-RAB13 antibody [EPR14109(B)] - BSA and Azide free (AB250269)

This data was developed using ab180936, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-RAB13 antibody [EPR14109(B)] (<a href='/en-us/products/primary-antibodies/rab13-antibody-epr14109b-ab180936'>ab180936</a>) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in 1%SDS Hot lysis method at 20 µg

Lane 3:

Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 15 µg

Lane 4:

Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared in 1%SDS Hot lysis method at 15 µg

Lane 5:

SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 15 µg

Lane 6:

SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate prepared in 1%SDS Hot lysis method 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: 23 kDa

Observed band size: 23 kDa

false