Human RAB8A knockout HeLa cell lysate (ab257195)

RAB8A KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 423 bp deletion in exon 1.

Be the first to review this product! Submit a review

Images

Western blot - Human RAB8A knockout HeLa cell lysate (AB257195), expandable thumbnail

Key facts

Cell type: HeLa
Species or organism: Human
Tissue: Cervix
Knockout validation: Sanger Sequencing, Western blot
Mutation description: Knockout achieved by using CRISPR/Cas9, Homozygous: 423 bp deletion in exon 1.

Alternative names

AA409338, MEL, MGC124948, Mel transforming oncogene, Mel transforming oncogene (RAB8 homolog), Mel transforming oncogene (derived from cell line NK14), Mel transforming oncogene (derived from cell line NK14) RAB8 homolog, Oncogene c-mel, RAB8, RAB8A member RAS oncogene family, RAB8A_HUMAN, Ras associated protein RAB8, Ras-related protein Rab-8A

What's included?

1 Kit

Components

Human RAB8A knockout HeLa cell lysate

1 x 100 µg

Human wild-type HeLa cell lysate

1 x 100 µg

Recommended products

RAB8A KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 423 bp deletion in exon 1.

Key facts

Cell type: HeLa
Species or organism: Human
Tissue: Cervix
Knockout validation: Sanger Sequencing, Western blot
Mutation description: Knockout achieved by using CRISPR/Cas9, Homozygous: 423 bp deletion in exon 1.
Disease: Adenocarcinoma
Concentration

Properties

Gene name: RAB8A
Gene editing type: Knockout
Gene editing method: CRISPR technology
Knockout validation: Sanger Sequencing, Western blot
Zygosity: Homozygous

Quality control

STR analysis: CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level: EU: 2 US: 2
Adherent/suspension: Adherent
Gender: Female

Storage

Shipped at conditions: Ambient - Can Ship with Ice
Appropriate short-term storage conditions: -20°C
Appropriate long-term storage conditions: -20°C

Notes

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

RAB8A is a small GTPase also referred to as Ras-related protein Rab-8A involved in intracellular membrane trafficking. It plays a critical role in the regulation of exocytosis the process by which cells release substances to the extracellular space. RAB8A mainly functions by cycling between an inactive GDP-bound form and an active GTP-bound form facilitating vesicle budding transport and fusion with target membranes. This protein is expressed in various tissues including the brain and pancreas where it assists in important cellular functions. RAB8A has a molecular mass of approximately 24 kDa.

Biological function summary

The small GTPase RAB8A contributes to the biosynthetic transport from the trans-Golgi network to the plasma membrane. It performs this activity in coordination with other proteins as part of a larger complex. RAB8A also plays an essential role in cilia formation and maintenance affecting cell polarity and signaling functions. The protein's ability to regulate vesicle movement makes it important for cellular homeostasis and communication.

Pathways

The regulatory function of RAB8A is significant in the ciliary membrane trafficking pathway and the insulin signaling pathway. Within these pathways RAB8A interacts closely with other proteins like Rab11 and Rabin8 which help support its roles in directing vesicle transport. These interactions are essential for proper cellular signaling and the maintenance of polarized cellular structures further supporting the function of various cellular pathways.

Associated diseases and disorders

The dysfunction of RAB8A has associations with certain ciliopathies notably Bardet-Biedl syndrome. This condition involves abnormalities in cilia structure and function leading to symptoms like kidney disease vision impairment and obesity. Moreover disruptions in RAB8A have connections with insulin resistance implicating it in metabolic disorders. In the context of these diseases proteins such as BBS proteins and Rab11 play a role in mediating the effects of dysfunctional RAB8A-related pathways.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

Western blot - Human RAB8A knockout HeLa cell lysate (ab257195)
Lane 1: Wild-type HeLa cell lysate (20 μg)
Lane 2: RAB8A knockout HeLa cell lysate (20 μg)
Lane 3: HCT116 cell lysate (20 μg)
Lane 4: Human brain tissue lysate (20 μg)
Lanes 1-4: Merged signal (red and green). Green - Anti-RAB8A antibody [MJF-R22-79-3] ab241061 observed at 24 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-RAB8A antibody [MJF-R22-79-3] ab241061 Anti-RAB8A antibody [MJF-R22-79-3] was shown to specifically react with RAB8A in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human RAB8A knockout HeLa cell line ab264993 (knockout cell lysate ab257195) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. ab Anti-RAB8A antibody [MJF-R22-79-3] ab241061 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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 [MJF-R22-79-3] (Anti-RAB8A antibody [MJF-R22-79-3] ab241061) 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
Performed under reducing conditions.
Predicted band size: 24 kDa
Observed band size: 24 kDa
Western blot - Human RAB8A knockout HeLa cell lysate (ab257195)
Lane 1: Wild-type HeLa cell lysate (20 μg)
Lane 2: RAB8A knockout HeLa cell lysate (20 μg)
Lane 3: HCT116 cell lysate (20 μg)
Lane 4: Human brain tissue lysate (20 μg)
Lanes 1-4: Merged signal (red and green). Green - Anti-RAB8A antibody [EPR14873] ab188574 observed at 24 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-RAB8A antibody [EPR14873] 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 Human RAB8A knockout HeLa cell line ab264993 (knockout cell lysate ab257195) was used. Wild-type and RAB8A knockout samples were subjected to SDS-PAGE. Anti-RAB8A antibody [EPR14873] ab188574 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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] (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
Performed under reducing conditions.
Predicted band size: 24 kDa
Observed band size: 24 kDa
Western blot - Human RAB8A knockout HeLa cell lysate (ab257195)
Anti-RAB8A antibody [EPR14873] 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 Human RAB8A knockout HeLa 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 Anti-RAB8A antibody [EPR14873] 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] (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 (Human RAB8A knockout HeLa cell line ab264993)
Western blot - Human RAB8A knockout HeLa cell lysate (ab257195)
Anti-RAB8A antibody [MJF-R22-79-3] ab241061 was shown to react with RAB8A in wild-type HeLa cells in Western blot with loss of signal observed in RAB8A knockout cell line Human RAB8A knockout HeLa 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 Anti-RAB8A antibody [MJF-R22-79-3] ab241061 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 [MJF-R22-79-3] (Anti-RAB8A antibody [MJF-R22-79-3] ab241061) 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 (Human RAB8A knockout HeLa cell line ab264993)
Western blot - Human RAB8A knockout HeLa cell lysate (ab257195)
Anti-RAB8A antibody [MJF-R22] ab237702 was shown to react with RAB8A in wild-type HeLa cells in Western blot with loss of signal observed in RAB8A knockout cell line Human RAB8A knockout HeLa 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 Anti-RAB8A antibody [MJF-R22] ab237702 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 [MJF-R22] (Anti-RAB8A antibody [MJF-R22] ab237702) 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 (Human RAB8A knockout HeLa cell line ab264993)
Immunocytochemistry/ Immunofluorescence - Human RAB8A knockout HeLa cell lysate (ab257195)
Anti-RAB8A antibody [MJF-R22] ab237702 was shown to react with RAB8A in wild-type HeLa cells in immunocytochemistry with loss of signal observed in RAB8A knockout cell line Human RAB8A knockout HeLa cell line ab264993. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with Anti-RAB8A antibody [MJF-R22] ab237702 at 1/250 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 ug/ml.

Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

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.
Sanger Sequencing - Human RAB8A knockout HeLa cell lysate (ab257195)
Homozygous: 423 bp deletion in exon 1
Immunocytochemistry/ Immunofluorescence - Human RAB8A knockout HeLa cell lysate (ab257195)
Anti-RAB8A antibody [MJF-R22-79-3] ab241061 was shown to react with RAB8A in wild-type HeLa cells in immunocytochemistry with loss of signal observed in RAB8A knockout cell line Human RAB8A knockout HeLa cell line ab264993. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with Anti-RAB8A antibody [MJF-R22-79-3] ab241061 at 1/600 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 ug/ml.

Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

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.