Overview

  • Product name
  • Description
    Rabbit polyclonal to Flotillin 1
  • Host species
    Rabbit
  • Specificity
    This target has been reported to show both cytoplasmic and nuclear localisation (PMID: 15713644).
  • Tested applications
    Suitable for: 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.

    Read Abcam's proprietary immunogen policy (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-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

Images

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

    Lanes 1 - 4: Merged signal (red and green). Green - ab41927 observed at 47 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab41927 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. ab41927 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml 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.

  • 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

  • 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 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : 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 (1/100) 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

  • 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.

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

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : 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

References

This product has been referenced in:
  • Xie JC  et al. Hypoxia increases amyloid-ß level in exosomes by enhancing the interaction between CD147 and Hook1. Am J Transl Res 10:150-163 (2018). Read more (PubMed: 29423001) »
  • Real JM  et al. Exosomes from patients with septic shock convey miRNAs related to inflammation and cell cycle regulation: new signaling pathways in sepsis? Crit Care 22:68 (2018). Read more (PubMed: 29540208) »
See all 23 Publications for this product

Customer reviews and Q&As

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1-10 of 10 Abreviews

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

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

Prof. Ritva Tikkanen

Verified customer

Submitted Jan 11 2013

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

Dr. Mahesh Shivananjappa

Verified customer

Submitted May 24 2012

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

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

Abcam user community

Verified customer

Submitted Aug 23 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (hippocampus)
Loading amount
25 µg
Specification
hippocampus
Gel Running Conditions
Reduced Denaturing (12% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 11 2011

Abcam has not validated the combination of species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Platelets)
Specification
Platelets
Preparation
Cell harvesting/tissue preparation method: PL were isolated spinning Platelet rich plasma on Histopaque
Sample buffer: PBS
Fixation
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton-X100 in 2% BSA for 30min
Gating Strategy
Platelets

Dr. Mahesh Shivananjappa

Verified customer

Submitted Jan 11 2011

Application
Immunocytochemistry
Sample
Human Cultured Cells (U937)
Specification
U937
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 27°C

Dr. Mahesh Shivananjappa

Verified customer

Submitted Oct 06 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (human (anal cancer), mouse (brain), rat (spleen))
Specification
human (anal cancer), mouse (brain), rat (spleen)
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization
No
Blocking step
sequential peroxidase and protein block (prediluted) as blocking agent for 20 minute(s) · Concentration: 100%

Mr. Antibody Solutions

Verified customer

Submitted Apr 28 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Methanol
Permeabilization
No

Dr. Kirk Mcmanus

Verified customer

Submitted Feb 18 2008

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

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