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AB265325

Human ARL8B knockout HeLa cell line

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ARL8B KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View Alternative Names

ADP ribosylation factor like 10C, ADP ribosylation factor like 8B, ADP-ribosylation factor-like protein 10C, ADP-ribosylation factor-like protein 8B, ARL10C, ARL8B_HUMAN, FLJ10702, Gie1, Novel small G protein indispensable for equal chromosome segregation 1

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Sanger Sequencing - Human ARL8B knockout HeLa cell line (AB265325)
  • Sanger seq

Unknown

Sanger Sequencing - Human ARL8B knockout HeLa cell line (AB265325)

Homozygous : 1 bp insertion in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1

Disease

Adenocarcinoma

Product details

We will provide viable cells that proliferate on revival.

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

What's included?

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Properties and storage information

Gene name
ARL8B
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

Supplementary information

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

ARL8B also known as ADP-ribosylation factor-like protein 8B is a small GTPase with an approximate molecular mass of 22 kDa. It mainly localizes to the lysosomal membrane. This protein plays an important role in the regulation of intracellular trafficking and lysosome positioning. ARL8B is expressed across many tissues suggesting its involvement in broad cellular functions.
Biological function summary

ARL8B helps manage the proper function of lysosomes by facilitating their movement and fusion with other cellular compartments. It forms a complex with other proteins such as PLEKHM2 and HOPS which are necessary for tethering events during lysosomal trafficking. This protein contributes to cellular processes such as autophagy and antigen presentation influencing the cell's ability to maintain homeostasis and effectively respond to external stimuli.

Pathways

ARL8B participates in the autophagy pathway and plays a part in the lysosome-related processes. It works closely with proteins like the kinesin motor proteins which drive the movement of lysosomes along microtubules. Through these interactions ARL8B coordinates with other autophagy-related proteins to ensure efficient degradation of cellular debris and recycling of components which are important for cellular health and function.

ARL8B has links to neurodegenerative diseases and immune system dysfunctions. Dysregulation of lysosomal positioning and movement caused by abnormal ARL8B function can lead to conditions such as Charcot-Marie-Tooth disease and impaired immune responses. Furthermore ARL8B interacts with proteins like Rab7a which can have implications in disease pathways emphasizing its significant role in maintaining cellular balance.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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