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AB267276

Human FLOT1 (Flotillin 1) knockout HEK-293T cell line

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FLOT1 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: 8 bp deletion in exon 10. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (AB267276)
  • WB

Lab

Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (AB267276)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : FLOT1 knockout HEK293T cell lysate (20 ug)
Lane 3 : HAP1 cell lysate (20 ug)

ab133497 was shown to specifically react with Flotillin 1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267276 (knockout cell lysate ab257109) was used. Wild-type and Flotillin 1 knockout samples were subjected to SDS-PAGE. ab133497 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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 1 antibody [EPR6041] (<a href='/en-us/products/primary-antibodies/flotillin-1-antibody-epr6041-ab133497'>ab133497</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-flot1-flotillin-1-knockout-hek-293t-cell-lysate-ab257109'>ab257109</a>) at 20 µg

Lane 3:

HAP1 cell lysate at 20 µg

Secondary

Lanes 1 - 3:

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 1/20000 dilution

Lanes 1 - 3:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 47 kDa

Observed band size: 50 kDa,36 kDa

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Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (AB267276)
  • WB

Unknown

Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (AB267276)

Lanes 1-4 : Merged signal (red and green). Green - ab78178 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab78178 Anti-Flotillin 1 antibody [EP446Y] was shown to specifically react with Flotillin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267276 (knockout cell lysate ab257109) was used. Wild-type and Flotillin knockout samples were subjected to SDS-PAGE. ab78178 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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 1 antibody [EP446Y] (<a href='/en-us/products/primary-antibodies/flotillin-1-antibody-ep446y-ab78178'>ab78178</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

FLOT1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (ab267276)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

HAP1 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: 47 kDa

Observed band size: 50 kDa

false

Cell Culture - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (AB267276)
  • Cell Culture

Unknown

Cell Culture - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (AB267276)

Representative images of FLOT1 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 an EVOS M5000 microscope.

Sanger Sequencing - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (AB267276)
  • Sanger seq

Unknown

Sanger Sequencing - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (AB267276)

Homozygous : 8 bp deletion in exon10

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: 8 bp deletion in exon 10

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Complementary Products

Primary Antibodies

AB133497

Anti-Flotillin 1 antibody [EPR6041]

RabMAb|Recombinant|KO Validated|20ul selling size

Flow Cytometry (Intracellular) - Anti-Flotillin 1 antibody [EPR6041] (AB133497)

  • 5
  • 6
Primary Antibodies

AB92536

Anti-MMP2 antibody [EPR1184]

RabMAb|Recombinant

Western blot - Anti-MMP2 antibody [EPR1184] (AB92536)

  • 4
  • 6
Primary Antibodies

AB26

Anti-p53 antibody [PAb 240]

KO Validated|Lab Essentials

Western blot - Anti-p53 antibody [PAb 240] (AB26)

  • 4
  • 56
Primary Antibodies

AB76003

Anti-MMP9 antibody [EP1254]

RabMAb|Recombinant|KO Validated

Western blot - Anti-MMP9 antibody [EP1254] (AB76003)

  • 4
  • 12
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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
FLOT1
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 1 also known as flotillin-1 or flot-1 functions as a major scaffolding protein in lipid rafts which are specialized membrane microdomains. It has a molecular weight of approximately 47 kDa facilitating protein interactions within the cell membrane. Flotillin 1 localizes primarily to non-caveolar lipid rafts and is expressed widely across various tissue types. High expression levels occur in brain tissue contributing to its recognition as an important element within neuronal membranes.
Biological function summary

Flotillin-1 participates in vesicle trafficking and endocytosis by forming a complex with its counterpart flotillin-2. This complex stabilizes membrane microdomains and assists in signaling pathways. Flotillin-1 plays a significant role in cell adhesion migration and cytoskeleton organization by interacting with actin and other cytoskeletal proteins. By doing so it influences cellular processes related to membrane shaping and intracellular transport.

Pathways

Flotillin-1 integrates into the MAPK signaling pathway and the Wnt signaling pathway which are critical for cell differentiation and proliferation. It interacts with several proteins including caveolin-1 that have known roles in these pathways. These interactions help modulate signaling cascades providing necessary cellular responses to various extracellular stimuli.

Flotillin-1 has associations with Alzheimer's disease and various cancers. In Alzheimer's disease it interacts with amyloid precursor protein influencing amyloid-beta production and deposition. In cancer altered expression levels of flotillin-1 affect tumor cell adhesion and migration with its interactions impacting the expression of proteins like E-cadherin which is associated with cancer metastasis and progression. Understanding these interactions offers potential insights for developing therapeutic strategies.
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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 FLOT1
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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