Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (AB221780)

Intracellular flow cytometrical analysis of HeLa cells labeling RAB8A with ab188574 at 1/260 (pink) compared to a Isotype control (green). FITC-conjugated goat-anti-rabbit secondary antibody at 1/150 were used for the analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab188574).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (AB221780)

Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling RAB8A with ab188574 at 1/500. A prediluted HRP Polymer for Rabbit IgG was used as the secondary antibody. Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab188574).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (AB221780)

Immunofluorescent analysis of 4% paraformaldehyde-fixed HepG2 cells labelling RAB8A with ab188574 at 1/500 followed by Goat anti rabbit IgG (Alexa Fluor^® 488) and DAPI staining.

This data was developed using the same antibody clone in a different buffer formulation containing PBS BSA glycerol and sodium azide (ab188574).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (AB221780)

This IHC data was generated using the same anti-RAB8A antibody clone EPR14873 in a different buffer formulation (cat# ab188574).

Immunohistochemical analysis of paraffin-embedded Human adenocarcinoma of colon tissue labelling RAB8A with ab188574 at 1/500. A prediluted HRP Polymer for Rabbit IgG was used as the secondary antibody. Counter stained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (AB221780)

This data was developed using the same antibody clone in a different buffer formulation (ab188574).

Lanes 1-4 : Merged signal (red and green). Green - ab188574 observed at 24 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab188574 Anti-RAB8A antibody [EPR14873] was shown to specifically react with RAB8A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264993 (knockout cell lysate ab257195) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab188574 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-RAB8A antibody [EPR14873] (<a href='/en-us/products/primary-antibodies/rab8a-antibody-epr14873-ab188574'>ab188574</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

RAB8A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human RAB8A knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-rab8a-knockout-hela-cell-line-ab264993'>ab264993</a>)

Lane 3:

HCT116 cell lysate at 20 µg

Lane 4:

Human brain tissue lysate at 20 µg

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-RAB8A antibody [EPR14873] - Low endotoxin, Azide free (AB221780)

This WB data was generated using the same anti-RAB8A antibody clone, EPR14873, in a different buffer formulation (cat# ab188574).

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : RAB8A knockout HAP1 cell lysate (20 μg)
Lane 3 : HCT116 cell lysate (20 μg)
Lane 4 : NIH/3T3 cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab188574 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab188574 was shown to specifically react with RAB8A when RAB8A knockout samples were used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab188574 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-RAB8A antibody [EPR14873] (<a href='/en-us/products/primary-antibodies/rab8a-antibody-epr14873-ab188574'>ab188574</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

RAB8A knockout HAP1 cell lysate at 20 µg

Lane 3:

HCT116 cell lysate at 20 µg

Lane 4:

NIH/3T3 cell lysate at 20 µg

Predicted band size: 24 kDa

false