Anti-RAB39B antibody [EPR9651]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-RAB39B antibody [EPR9651] (AB154826)

Intracellular Flow Cytometry analysis of U87-MG cells labelling RAB39B with unpurified ab154826 (red) at 1/9. Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RAB39B antibody [EPR9651] (AB154826)

Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y cells labelling RAB39B with purified ab154826 at 1/200 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 555-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RAB39B antibody [EPR9651] (AB154826)

Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y cells labelling RAB39B with unpurified ab154826 at 1/10 (red). Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 555-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-RAB39B antibody [EPR9651] (AB154826)

Intracellular Flow Cytometry analysis of U87-MG cells labelling RAB39B with purified ab154826 (red) at 1/210. Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-RAB39B antibody [EPR9651] (AB154826)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling RAB39B with ab154826 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 μg/mL) (Green). Confocal image showing positive staining in mouse primary neuron cell.Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (4 μg/mL) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (2 μg/mL) (Red). The nuclear counterstain was DAPI (Blue).

• WB

Lab

Western blot - Anti-RAB39B antibody [EPR9651] (AB154826)

Western blot : Rabbit Monoclonal[EPR9651] to RAB39B ab154826 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 25 kDa in Wild-type U-87 MG UNBOILED cell lysates with no signal observed at this size in RAB39B knockout U-87 MG UNBOILED 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 & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-RAB39B antibody [EPR9651] (ab154826) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG UNBOILED at 20 µg

Lane 2:

Western blot - Human RAB39B knockout U-87 MG cell line (ab306767) at 20 µg

Lane 3:

SH-SY5Y UNBOILED at 20 µg

Lane 4:

Human Brain UNBOILED at 20 µg

Lane 5:

K562 UNBOILED at 20 µg

Secondary

All lanes:

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

Predicted band size: 25 kDa

Observed band size: 25 kDa

false

• WB

Lab

Western blot - Anti-RAB39B antibody [EPR9651] (AB154826)

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RAB39B antibody [EPR9651] (ab154826) at 1/100 dilution

Lane 1:

Human fetal brain tissue lysate at 10 µg

Lane 2:

U87-MG cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 25 kDa

Observed band size: 25 kDa

false

• WB

Lab

Western blot - Anti-RAB39B antibody [EPR9651] (AB154826)

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RAB39B antibody [EPR9651] (ab154826) at 1/2000 dilution

Lane 1:

Human fetal brain tissue lysate at 10 µg

Lane 2:

U87-MG cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 25 kDa

Observed band size: 25 kDa

false

• WB

Unknown

Western blot - Anti-RAB39B antibody [EPR9651] (AB154826)

All lanes:

Western blot - Anti-RAB39B antibody [EPR9651] (ab154826) at 1/1000 dilution

Lane 1:

Fetal brain lysate at 10 µg

Lane 2:

U87-MG cell lysate at 10 µg

Lane 3:

SH-SY5Y cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 25 kDa

false