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AB266847

Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line

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FLOT2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 4. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View 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)

3 Images
Western blot - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (AB266847)
  • WB

Lab

Western blot - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (AB266847)

Lanes 1-4 : Merged signal (red and green). Green - ab181988 observed at 47 kDa. Red - loading control ab8245 observed at 36 kDa.

ab181988 Anti-Flotillin 2/ESA antibody [EPR14128(B)] was shown to specifically react with Flotillin 2/ESA in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266847 (knockout cell lysate ab257442) was used. Wild-type and Flotillin 2/ESA knockout samples were subjected to SDS-PAGE. ab181988 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (<a href='/en-us/products/primary-antibodies/flotillin-2-esa-antibody-epr14128b-ab181988'>ab181988</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

FLOT2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (ab266847)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

A549 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 27 kDa,47 kDa

Observed band size: 27 kDa,47 kDa

false

Sanger Sequencing - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (AB266847)
  • Sanger seq

Unknown

Sanger Sequencing - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (AB266847)

Homozygous : 5 bp deletion in exon4

Cell Culture - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (AB266847)
  • Cell Culture

Lab

Cell Culture - Human FLOT2 (Flotillin 2/ESA) knockout HEK-293T cell line (AB266847)

Representative images FLOT2 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

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.

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What's included?

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Properties and storage information

Gene name
FLOT2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Supplementary information

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

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.

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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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Target data

See full target information FLOT2
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Complementary Products

Primary Antibodies

AB181988

Anti-Flotillin 2/ESA antibody [EPR14128(B)]

RabMAb|Recombinant|KO Validated

Immunoprecipitation - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (AB181988)

  • 5
  • 2
Primary Antibodies

AB125011

Anti-TSG101 antibody [EPR7130(B)]

BOND RX™ Validated|RabMAb|Recombinant

Immunohistochemistry - Anti-TSG101 antibody [EPR7130(B)] (AB125011)

  • 4
  • 10
Primary Antibodies

AB238126

Anti-GRP94 antibody [EPR22847-50]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] (AB238126)

  • 0
  • 0
Primary Antibodies

AB178677

Anti-Annexin-2/ANXA2 antibody [EPR13052(B)]

RabMAb|Recombinant

Flow Cytometry (Intracellular) - Anti-Annexin-2/ANXA2 antibody [EPR13052(B)] (AB178677)

  • 0
  • 0

Product promise

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