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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microfilaments Actin etc Actin Binding Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Flotillin 1 antibody [EPR6041] (ab133497)

  • Datasheet
  • SDS
Submit a review Submit a question References (20)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Immunocytochemistry/ Immunofluorescence - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Immunoprecipitation - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Flow Cytometry - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
  • Anti-Flotillin 1 antibody [EPR6041] (ab133497)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6041] to Flotillin 1
  • Suitable for: Flow Cyt, WB, IHC-P, IP, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-Flotillin 1 antibody [EPR6041]
    See all Flotillin 1 primary antibodies
  • Description

    Rabbit monoclonal [EPR6041] to Flotillin 1
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Mouse
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human Flotillin 1 aa 400-500 (C terminal). The exact sequence is proprietary.
    Database link: O75955

  • Positive control

    • WB: HEK293T, HAP1, K562, A431, Jurkat, and HeLa cell lysates; Mouse brain and kidney lysates; Rat kidney lysate. IP: K562 cell lysate. IHC-P: Human cervical carcinoma tissue. Flow Cyt: Jurkat cells. ICC/IF: Jurkat cells.
  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR6041
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Binding Proteins
    • Signal Transduction
    • Protein Trafficking
    • Vesicle Transport
    • Coat Proteins
    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Other cell division antibodies

Associated products

  • Alternative Versions

    • APC Anti-Flotillin 1 antibody [EPR6041] (ab225164)
    • PE Anti-Flotillin 1 antibody [EPR6041] (ab225165)
    • Anti-Flotillin 1 antibody [EPR6041] - BSA and Azide free (ab232405)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human FLOT1 (Flotillin 1) knockout HEK-293T cell line (ab267276)
  • KO cell lysates

    • Human FLOT1 (Flotillin 1) knockout HEK-293T cell lysate (ab257109)
  • Positive Controls

    • HeLa membrane extract lysate (ab29547)
    • Jurkat membrane extract lysate (ab30130)
    • A-431 membrane extract lysate (ab30134)
  • Recombinant Protein

    • Recombinant Human Flotillin 1 protein (ab153058)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133497 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Mouse
Human
Application Abreviews Notes
Flow Cyt
1/50.
WB
1/10000 - 1/50000. Detects a band of approximately 48 kDa (predicted molecular weight: 47 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

IP
1/10 - 1/100.
ICC/IF
1/500.
Notes
Flow Cyt
1/50.
WB
1/10000 - 1/50000. Detects a band of approximately 48 kDa (predicted molecular weight: 47 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

IP
1/10 - 1/100.
ICC/IF
1/500.

Target

  • Function

    May act as a scaffolding protein within caveolar membranes, functionally participating in formation of caveolae or caveolae-like vesicles.
  • Sequence similarities

    Belongs to the band 7/mec-2 family. Flotillin subfamily.
  • Cellular localization

    Cell membrane. Membrane > caveola. Melanosome. Endosome. Membrane-associated protein of caveolae. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Target information above from: UniProt accession O75955 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10211 Human
    • Entrez Gene: 14251 Mouse
    • Entrez Gene: 64665 Rat
    • Omim: 606998 Human
    • SwissProt: O75955 Human
    • SwissProt: O08917 Mouse
    • SwissProt: Q9Z1E1 Rat
    • Unigene: 179986 Human
    • Unigene: 2931 Mouse
    • Unigene: 9567 Rat
    see all
  • Alternative names

    • FLOT 1 antibody
    • FLOT1 antibody
    • FLOT1_HUMAN antibody
    • Flotillin-1 antibody
    • Flotillin1 antibody
    • Integral membrane component of caveolae antibody
    • Reggie 2 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab133497 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Immunocytochemistry/ Immunofluorescence - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Immunocytochemistry/ Immunofluorescence - Anti-Flotillin 1 antibody [EPR6041] (ab133497)

    Immunofluorescence staining of Jurkat cells with purified ab133497 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab133497 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    All lanes : Anti-Flotillin 1 antibody [EPR6041] (ab133497) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : FLOT1 knockout HEK293T cell lysate
    Lane 3 : HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?



    Lanes 1-3: Merged signal (red and green). Green - ab133497 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab133497 Anti-Flotillin 1 antibody [EPR6041] 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 4°C 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.

  • Immunoprecipitation - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Immunoprecipitation - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    ab133497 (purified) at 1/60 immunoprecipitating Flotillin 1 in 10 μg K562 (Lanes 1 and 2, observed at 48 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
  • Flow Cytometry - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Flow Cytometry - Anti-Flotillin 1 antibody [EPR6041] (ab133497)

    Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling Flotillin 1 with purified ab133497 at 1/50 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Flotillin 1 antibody [EPR6041] (ab133497)

    Immunohistochemical analysis of paraffin embedded Human colon tissue labelling Flotillin 1 using unpurified ab133497 at a dilution of 1/100.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Flotillin 1 knockout HAP1 whole cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green - ab133497 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab133497 was shown to specifically react with Flotillin 1 in wild-type HAP1 cells as signal was lost in Flotillin 1 knockout cells. Wild-type and Flotillin 1 knockout samples were subjected to SDS-PAGE. Ab133497 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    All lanes : Anti-Flotillin 1 antibody [EPR6041] (ab133497) at 1/10000 dilution (purified)

    Lane 1 : K562 cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 47 kDa
    Observed band size: 48 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    All lanes : Anti-Flotillin 1 antibody [EPR6041] (ab133497) at 1/10000 dilution (purified)

    Lane 1 : mouse brain lysate
    Lane 2 : mouse kidney lysate
    Lane 3 : rat kidney lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L)

    Predicted band size: 47 kDa
    Observed band size: 48 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Western blot - Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    All lanes : Anti-Flotillin 1 antibody [EPR6041] (ab133497) at 1/10000 dilution

    Lane 1 : K562 cell lysate
    Lane 2 : A431 cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 47 kDa
    Observed band size: 48 kDa why is the actual band size different from the predicted?

  • Anti-Flotillin 1 antibody [EPR6041] (ab133497)
    Anti-Flotillin 1 antibody [EPR6041] (ab133497)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (20)

    Publishing research using ab133497? Please let us know so that we can cite the reference in this datasheet.

    ab133497 has been referenced in 20 publications.

    • Zhu W  et al. CD41-deficient exosomes from non-traumatic femoral head necrosis tissues impair osteogenic differentiation and migration of mesenchymal stem cells. Cell Death Dis 11:293 (2020). PubMed: 32341357
    • Maas SLN  et al. Glioblastoma hijacks microglial gene expression to support tumor growth. J Neuroinflammation 17:120 (2020). PubMed: 32299465
    • Gao L  et al. Cardio-renal Exosomes in Myocardial Infarction Serum Regulate Proangiogenic Paracrine Signaling in Adipose Mesenchymal Stem Cells. Theranostics 10:1060-1073 (2020). PubMed: 31938051
    • Chang YT  et al. Pancreatic cancer-derived small extracellular vesical Ezrin regulates macrophage polarization and promotes metastasis. Am J Cancer Res 10:12-37 (2020). PubMed: 32064151
    • Lv Y  et al. A recombinant measles virus vaccine strain rMV-Hu191 has oncolytic effect against human gastric cancer by inducing apoptotic cell death requiring integrity of lipid raft microdomains. Cancer Lett 460:108-118 (2019). PubMed: 31226409
    View all Publications for this product

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