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FLOT2 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: 5 bp deletion in exon 4.

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Images

Western blot - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (AB266847), expandable thumbnail
  • Cell Culture - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (AB266847), expandable thumbnail
  • Sanger Sequencing - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (AB266847), expandable thumbnail

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

Knockout validation

Sanger Sequencing, Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 4

Alternative names

ECS-1, ESA, ESA1, Epidermal surface antigen, Epidermal surface antigen 1, FLOT2_HUMAN, Flotillin 2 (epidermal surface antigen 1), Flotillin-2, M17S1, Membrane component chromosome 17 surface marker 1, Membrane component chromosome 17 surface marker 1 homolog, Reggie-1, Reggie-2, membrane component, chromosome 17, surface marker 1 (35kD protein identified by monoclonal antibody ECS-1)

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FLOT2 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: 5 bp deletion in exon 4.

Key facts

Cell type

HEK-293T

Form

Liquid

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 4

Concentration
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Properties

Gene name

FLOT2

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 HEK293T cell line (Human wild-type HEK-293T 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.

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

Flotillin 2 also known as flotillin-2 or ESA is a protein weighing approximately 48 kDa and is a part of the flotillin family. It is expressed in various human tissues with high presence in the brain heart and skeletal muscles. This protein is an integral component of the lipid raft microdomains in cell membranes where it assists in cellular processes such as signal transduction and lipid trafficking. Its membrane association depends on N-terminal hydrophobic sequences which contribute to its distribution on the cell surface.

Biological function summary

Flotillins serve as scaffolding proteins in cellular signaling and play a part in the regulation of endocytosis. Flotillin 2 often forms a hetero-oligomeric complex with Flotillin 1 facilitating its role in organizing membrane microdomains. This interaction supports cellular activities like molecular transport and signal relay. Flotillin 2 contributes to dynamic cellular structures and has been suggested to influence actin cytoskeleton assembly giving it a role in cellular morphology and motility.

Pathways

Flotillin 2 associates with important cell signaling pathways notably the MAPK and Wnt signaling pathways. In MAPK pathways flotillin 2 influences cell proliferation and differentiation by modulating interaction with signaling proteins like Ras and ERK. In the context of the Wnt signaling pathway it impacts cellular communication influencing gene expression and development processes by engaging with Dishevelled proteins. These pathways highlight its critical part in processes like cellular growth and communication.

Associated diseases and disorders

Flotillin 2 exhibits connections regarding metabolic and neurodegenerative conditions. Flotillin 2 upregulation appears linked with metabolic syndromes associating with insulin signaling disruptions. Furthermore alterations in its expression relate to neurodegenerative diseases like Alzheimer's where it impacts amyloid precursor protein processing and tau protein pathology. These associations position flotillin 2 as a significant player in the understanding and potential treatment of these disorders.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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