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AB257109

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

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FLOT1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 8 bp deletion in exon10.

View Alternative Names

FLOT1_HUMAN, Flotillin-1, Integral membrane component of caveolae, Reggie-2

3 Images
Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell lysate (AB257109)
  • WB

Lab

Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell lysate (AB257109)

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:

FLOT1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-flot1-flotillin-1-knockout-hek-293t-cell-line-ab267276'>ab267276</a>)

Lane 3:

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

Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell lysate (AB257109)
  • WB

Unknown

Western blot - Human FLOT1 (Flotillin 1) knockout HEK-293T cell lysate (AB257109)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : FLOT1 knockout HEK293T cell lysate (20 ug)
Lane 3 : A431 cell lysate (20 ug)
Lane 4 : HAP1 cell lysate (20 ug)

ab78178 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. ab78178 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 [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 (<a href='/en-us/products/cell-lines/human-flot1-flotillin-1-knockout-hek-293t-cell-line-ab267276'>ab267276</a>)

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

Sanger Sequencing - Human FLOT1 (Flotillin 1) knockout HEK-293T cell lysate (AB257109)
  • Sanger seq

Unknown

Sanger Sequencing - Human FLOT1 (Flotillin 1) knockout HEK-293T cell lysate (AB257109)

Homozygous : 8 bp deletion in exon10

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 8 bp deletion in exon10.

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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

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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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Product promise

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