Anti-RAB8A antibody [EPR14873]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] (AB188574)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RAB8A antibody [EPR14873] (AB188574)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB8A antibody [EPR14873] (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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RAB8A antibody [EPR14873] (AB188574)

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.

• IP

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Immunoprecipitation - Anti-RAB8A antibody [EPR14873] (AB188574)

Immunoprecipitation of RAB8A in HeLa cells. Lysates were prepared and immunoprecipitation was performed using 2 ug of ab188574 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

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.

• WB

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Western blot - Anti-RAB8A antibody [EPR14873] (AB188574)

ab188574 was shown to react with RAB8A in wild-type HeLa cells in Western blot with loss of signal observed in RAB8A knockout cell line ab264993. Wild-type HeLa and RAB8A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab188574 overnight at 4 °C at a 1/1000 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-RAB8A antibody [EPR14873] (ab188574) at 1/1000 dilution

Lane 1:

Wild-type HeLa lysate at 30 µg

Lane 2:

RAB8A knock-out HeLa lysate at 30 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>)

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• WB

Supplier Data

Western blot - Anti-RAB8A antibody [EPR14873] (AB188574)

All lanes:

Western blot - Anti-RAB8A antibody [EPR14873] (ab188574) at 1/1000 dilution

All lanes:

HCT-116 lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Lab

Western blot - Anti-RAB8A antibody [EPR14873] (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] (ab188574) 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

Unknown

Western blot - Anti-RAB8A antibody [EPR14873] (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] (ab188574) 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 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] (AB188574)

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] (ab188574) 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

• WB

Supplier Data

Western blot - Anti-RAB8A antibody [EPR14873] (AB188574)

All lanes:

Western blot - Anti-RAB8A antibody [EPR14873] (ab188574) at 1/10000 dilution

Lane 1:

HepG2 lysate at 20 µg

Lane 2:

HeLa lysate at 20 µg

Lane 3:

293T lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false

• WB

Supplier Data

Western blot - Anti-RAB8A antibody [EPR14873] (AB188574)

All lanes:

Western blot - Anti-RAB8A antibody [EPR14873] (ab188574) at 1/1000 dilution

Lane 1:

C6 lysate at 10 µg

Lane 2:

RAW264.7 lysate at 10 µg

Lane 3:

PC-12 lysate at 10 µg

Lane 4:

NIH3T3 lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 24 kDa

Observed band size: 24 kDa

false