- Datasheet
- Specific References (21)
- Protocols
Overview
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Product nameAnti-Flotillin 1 antibody
See all Flotillin 1 primary antibodies -
DescriptionRabbit polyclonal to Flotillin 1
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Host speciesRabbit
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SpecificityThis target has been reported to show both cytoplasmic and nuclear localisation (PMID: 15713644).
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Tested applicationsSuitable for: IHC-P, ICC/IF, WB, IPmore details
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Species reactivityReacts with: Mouse, Rat, Dog, Human
Predicted to work with: a wide range of other species
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Flotillin 1.
(Peptide available as ab41926.)
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Positive control
- ab41927 gave a positive result in Human Lung tissue lysate, Human Brain tissue lysate and Hela whole cell lysate.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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
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FunctionMay act as a scaffolding protein within caveolar membranes, functionally participating in formation of caveolae or caveolae-like vesicles.
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Sequence similaritiesBelongs to the band 7/mec-2 family. Flotillin subfamily.
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Cellular localizationCell membrane. Membrane > caveola. Melanosome. Endosome. Membrane-associated protein of caveolae. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
- Information by UniProt
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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
see all -
Alternative names
- FLOT 1 antibody
- FLOT1 antibody
- FLOT1_HUMAN antibody
see all
Images
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Western blot - Anti-Flotillin 1 antibody (ab41927)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.
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Immunocytochemistry/ Immunofluorescence - Anti-Flotillin 1 antibody (ab41927)This image is part of an abreview submitted by Dr. Kirk McManusab41927 (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. -
Western blot - Anti-Flotillin 1 antibody (ab41927)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 -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Flotillin 1 antibody (ab41927)This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.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. -
Immunoprecipitation - Anti-Flotillin 1 antibody (ab41927)This image is courtesy of an Abreview submitted by Prof Ritva TikkanenFlotillin 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 -
Immunocytochemistry/ Immunofluorescence - Anti-Flotillin 1 antibody (ab41927)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.
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Western blot - Anti-Flotillin 1 antibody (ab41927)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
Protocols
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) »
Customer reviews and Q&As
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"