Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using ab316151, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling Rab11A with ab316151 at 1/50 (10.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in A431 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using ab316151, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A549 (human lung carcinoma epithelial cell) cells labelling Rab11A with ab316151 at 1/50 (10.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in A549 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using ab316151, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C6 ( rat glial tumor glial cell) cells labelling Rab11A with ab316151 at 1/50 (10.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in C6 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using ab316151, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling Rab11A with ab316151 at 1/50 (10.1 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in Neuro-2a cell line.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• WB

Supplier Data

Western blot - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using ab316151, the same antibody clone in a different buffer formulation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Rab11A antibody [EPR28202-94] (<a href='/ko/products/primary-antibodies/rab11a-antibody-epr28202-94-ab316151'>ab316151</a>) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

A549 transfected with siRNA specifically targeting Rab11A 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: 24 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using ab316151, the same antibody clone in a different buffer formulation.

The lanes 2 was developed using a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-Rab11A antibody [EPR28202-94] (<a href='/ko/products/primary-antibodies/rab11a-antibody-epr28202-94-ab316151'>ab316151</a>) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

C6 (rat glial tumor glial 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: 24 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using ab316151, the same antibody clone in a different buffer formulation.

The lanes 3-5 was developed using a high sensitivity ECL substrate.

Lanes 1-2 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000; Lanes 3-5 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000.

All lanes:

Western blot - Anti-Rab11A antibody [EPR28202-94] (<a href='/ko/products/primary-antibodies/rab11a-antibody-epr28202-94-ab316151'>ab316151</a>) at 1/1000 dilution

Lane 1:

Human brain tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat spleen tissue lysate at 20 µg

Secondary

Lanes 1 - 2:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 3 - 5:

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: 24 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab316151).

Western blot : Anti-Rab11A antibody [EPR28202-94] ab316151 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 24 kDa in Wild-type A549 cell lysates with no signal observed at this size in RAB11A 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 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-Rab11A antibody [EPR28202-94] (<a href='/ko/products/primary-antibodies/rab11a-antibody-epr28202-94-ab316151'>ab316151</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysates at 20 µg

Lane 2:

RAB11A knockout A549 cell lysates at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

HCT 116 cell lysates at 20 µg

Lane 5:

LNCaP cell lysates at 20 µg

Lane 6:

Jurkat cell lysates at 20 µg

Secondary

All lanes:

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

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Supplier Data

Western blot - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using ab316151, the same antibody clone in a different buffer formulation.

Rab11A has two splicing variants.

The identity of the lower MW band at approximately 18 kDa in lane 1-2 may represent an isoform of Rab11A (PMID : 14702039).

Exposure time : Lanes 1-2 : 70 seconds, Lane 3 : 180 seconds

All lanes:

Western blot - Anti-Rab11A antibody [EPR28202-94] (<a href='/ko/products/primary-antibodies/rab11a-antibody-epr28202-94-ab316151'>ab316151</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

EL4 (mouse lymphoma T lymphocyte) 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 1/20000 dilution

Observed band size: 24 kDa

false

• WB

Supplier Data

Western blot - Anti-Rab11A antibody [EPR28202-94] - BSA and Azide free (AB316152)

This data was developed using ab316151, the same antibody clone in a different buffer formulation.

This antibody does not cross-react with human Rab11B and Rab25.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Rab11A antibody [EPR28202-94] (<a href='/ko/products/primary-antibodies/rab11a-antibody-epr28202-94-ab316151'>ab316151</a>) at 1/1000 dilution

Lane 1:

His-tagged human Rab11A recombinant protein at 10 ng

Lane 2:

His-tagged human Rab11B recombinant protein at 10 ng

Lane 3:

His-tagged human Rab25 recombinant protein at 10 ng

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: 24 kDa

false

Exposure time: 180s