Overview

  • Product nameAnti-Flotillin 1 antibody
    See all Flotillin 1 primary antibodies
  • Description
    Rabbit polyclonal to Flotillin 1
  • SpecificityThis target has been reported to show both cytoplasmic and nuclear localisation (PMID: 15713644).
  • Tested applicationsIHC - Wholemount, IHC-P, ICC/IF, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
    Predicted to work with: a wide range of other species
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Flotillin 1.

    (Peptide available as ab41926.)

  • Positive control
    • ab41927 gave a positive result in Human Lung tissue lysate, Human Brain tissue lysate and Hela whole cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab41927 in the following tested applications.

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

Application Abreviews Notes
IHC - Wholemount Use at an assay dependent concentration.
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/200. Fix cells with methanol
WB Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).Can be blocked with Human Flotillin 1 peptide (ab41926).
IP Use at an assay dependent concentration.

Target

Anti-Flotillin 1 antibody images

  • All lanes : Anti-Flotillin 1 antibody (ab41927) at 1 µg/ml

    Lane 1 : Lung (Human) Tissue Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 47 kDa
    Observed band size : 47 kDa
  • ab41927 (1/200) staining Flotillin 1 in HeLa cells (green). Cells were fixed with Methanol and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • ab41927 (1/100) staining Flotilin 1 in paraffin embedded - Human (anal cancer), Mouse (brain) and Rat (spleen) tissues. Tissue sections underwent paraformaldehyde fixation, heat mediated antigen retrieval using Citrate Buffer pH 6.0 and were blocked using sequential peroxidase and protein block (prediluted). Please refer to abreview for further experimental details.

    See Abreview

  • Flotillin 1 immunoprecipitation from Human HeLa whole cell lysate using ab41927. 750 µg of cell lysate was incubated with primary antibody (1/500) and matrix (Protein A) and incubated for 16 hours at 4°C.

    Lane 1 : anti-Flotillin 2 IP (using non-Abcam antibody)
    Lane 2 : anti-Flotillin 2 IP (using ab41927)
    Lane 3 : Input (HeLa cell lysate)

    Western blot detection : mouse anti-Flotillin 1 antibody + HRP donkey anti-mouse polyclonal secondary antibody

    See Abreview

  • All lanes : Anti-Flotillin 1 antibody (ab41927) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 47 kDa
    Observed band size : 50 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 35 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes
  • ICC/IF image of ab41927 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab41927 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Flotillin 1 antibody (ab41927)

This product has been referenced in:
  • Burkhardt P  et al. Evolutionary Insights into Premetazoan Functions of the Neuronal Protein Homer. Mol Biol Evol N/A:N/A (2014). ICC/IF . Read more (PubMed: 24899667) »
  • Svensson KJ  et al. Exosome uptake depends on ERK1/2-heat shock protein 27 signaling and lipid Raft-mediated endocytosis negatively regulated by caveolin-1. J Biol Chem 288:17713-24 (2013). Read more (PubMed: 23653359) »

See all 11 Publications for this product

Product Wall

Application IHC - Wholemount
Sample Xenopus laevis Embryo (st.11.5, dorsal view)
Specification st.11.5, dorsal view
Username

Abcam user community

Verified customer

Submitted Mar 26 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (HeLa)
Total protein in input 750 µg
Specification HeLa
Immuno-precipitation step Protein A
Username

Prof. Ritva Tikkanen

Verified customer

Submitted Jan 11 2013

I am very pleased to hear you would like to test both ab41927 and ab96507 in immunoprecipitation. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for immunoprecipitation and in...

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Thank you for contacting us.

I'm sorry to hear that the antibody is not working anymore after the -80 degree storage.
As we discussed over the phone, both species are untested.

If you could test the antibody in human samples -...

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Application Immunocytochemistry
Sample Mouse Cell (RAW 264.7)
Specification RAW 264.7
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton-X100 in 2% BSA for 15min
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted May 24 2012

DISCOUNT CODE: ***
Expiration date: May 23rd, 2012

I am very pleased to hear you would like to accept our offer and test ab41927 in macaque. This code will give you 1 free primary antibody before the expiration date. To redeem this offer...

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Thank you for your enquiry. flotillin1 is a peripheral membrane protein that is associated with caveolae. As such I believe it will be a cytoplasmic (internal) epitope that is detected by this antibody. However you may wish to consult the caveolae lite...

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Thank you very much for your interest in ab41927. To our knowledge, ab41927 has not been tested in Flow cytometry. Therefore, I can offer a discount off a future purchase if you buy ab41927 now, test it in Flow cytometry and submit feedback to us in th...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Platelets)
Loading amount 25 µg
Specification Platelets
Gel Running Conditions Reduced Denaturing (4-15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Oct 31 2011

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Xenopus laevis Cell (embryos)
Specification embryos
Fixative Methanol
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 23 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"