Anti-Rab9 antibody [EPR13272] - Late Endosome Marker

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (AB179815)

Immunofluorescent staining of HepG2 cells labeling Rab9 with unpurified ab179815 at 1/50 dilution (red). DAPI nuclear staining (blue).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (AB179815)

Immunofluorescence staining of HepG2 cells with purified ab179815 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab179815 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

• WB

Supplier Data

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (AB179815)

All lanes:

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (ab179815) at 1/1000 dilution

Lane 1:

K562 cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

293T cell lysate at 10 µg

Lane 4:

HepG2 cell lysate at 10 µg

Predicted band size: 23 kDa

false

• WB

Lab

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (AB179815)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (ab179815) at 1/2000 dilution

Lane 1:

K562 cell lysate at 20 µg

Lane 2:

HeLa cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 23 kDa

Observed band size: 23 kDa

false

• WB

Lab

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (AB179815)

Lanes 1-4 : Merged signal (red and green). Green - ab179815 observed at 25 kDa. Red - loading control ab8245 observed at 36 kDa.

ab179815 Anti-Rab9 antibody [EPR13272] was shown to specifically react with Rab9 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265693 (knockout cell lysate ab257625) was used. Wild-type and Rab9 knockout samples were subjected to SDS-PAGE. ab179815 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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-Rab9 antibody [EPR13272] - Late Endosome Marker (ab179815) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

RAB9A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human RAB9A (Rab9) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-rab9a-rab9-knockout-hela-cell-line-ab265693'>ab265693</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

MDA-MB-231 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: 25 kDa

false

• WB

Lab

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (AB179815)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (ab179815) at 1/2000 dilution

All lanes:

rat kidney at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

false

• WB

Lab

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (AB179815)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Rab9 antibody [EPR13272] - Late Endosome Marker (ab179815) at 1/2000 dilution

All lanes:

mouse spleen at 20 µg

Secondary

All lanes:

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

Predicted band size: 23 kDa

Observed band size: 23 kDa

false