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AB266583

Human RAB14 knockout HEK-293T cell line

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RAB14 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp insertion in exon 4 and Insertion of the selection cassette in exon 4.

View Alternative Names

F protein binding protein 1, FBP, GTPase Rab14, RAB14 member RAS oncogene family, RAB14_HUMAN, RP11 165P4.4, Ras-related protein Rab-14, Small GTP binding protein RAB14, bA165P4.3

2 Images
Sanger Sequencing - Human RAB14 knockout HEK-293T cell line (AB266583)
  • Sanger seq

Unknown

Sanger Sequencing - Human RAB14 knockout HEK-293T cell line (AB266583)

Allele-1 : Insertion of the selection cassette in exon 4

Sanger Sequencing - Human RAB14 knockout HEK-293T cell line (AB266583)
  • Sanger seq

Unknown

Sanger Sequencing - Human RAB14 knockout HEK-293T cell line (AB266583)

Allele-2 : 2 bp insertion in exon 4.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 2 bp insertion in exon 4 and Insertion of the selection cassette in exon 4

Product details

Recommended control: Human wild-type HEK293T cell line (ab255449). 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.

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

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
RAB14
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
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.

RAB14 also known as Ras-related protein Rab-14 is part of the Rab family of small GTPases. It has a molecular mass of approximately 23 kDa and functions to regulate membrane trafficking between the Golgi apparatus and endosomes. RAB14 mediates vesicle movement and membrane fusion playing an important role in endocytic and secretory pathways. It is expressed in various tissues and cells including the brain heart and lungs suggesting its involvement in diverse physiological processes.
Biological function summary

RAB14 orchestrates intracellular transport processes important for cell homeostasis. It does not act alone but often forms complexes with other proteins like protein phosphatase 1 and interactions involving KIF3A and DVL. These interactions enable RAB14 to influence cargo sorting and organelle morphology which is essential for correct protein trafficking and recycling in the cells.

Pathways

Scientists have placed RAB14 in relation to the regulation of membrane trafficking and cell signaling. RAB14 influences the pathways of endocytosis and trans-Golgi network trafficking. It has associations with proteins such as RAB11 and RAB5 in these pathways. These proteins work in coordination to ensure effective sorting and distribution of intracellular materials affecting various cellular processes.

Mutations or dysregulation of RAB14 associate with cancer progression and metabolic disorders. Abnormal RAB14 activity influences tumorigenesis by affecting cell migration and invasion in part through interactions with proteins such as Rac1. Additionally RAB14 plays a role in insulin signaling pathways linking it to metabolic disorders related to insulin resistance and glucose homeostasis.

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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