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

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Images

Western blot - Human RAB8A knockout HeLa cell line (AB264993), expandable thumbnail
  • Western blot - Human RAB8A knockout HeLa cell line (AB264993), expandable thumbnail
  • Cell Culture - Human RAB8A knockout HeLa cell line (AB264993), expandable thumbnail
  • Sanger Sequencing - Human RAB8A knockout HeLa cell line (AB264993), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Human RAB8A knockout HeLa cell line (AB264993), expandable thumbnail

Key facts

Cell type
HeLa
Species or organism
Human
Tissue
Cervix
Form
Liquid
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

Recommended products

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

Key facts

Cell type
HeLa
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 423 bp deletion in exon 1
Antibiotic resistance
Puromycin 1µg/mL
Disease
Adenocarcinoma
Concentration
Loading...

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

Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

We will provide viable cells that proliferate on revival.

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.

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