製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Flotillin 1
  • 由来種
    Rabbit
  • 特異性
    This target has been reported to show both cytoplasmic and nuclear localisation (PMID: 15713644).
  • アプリケーション
    適用あり: IHC-P, ICC/IF, WB, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Dog, Human
    交差が予測される動物種: a wide range of other species
  • 免疫原

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

    (Peptide available as ab41926.)

  • ポジティブ・コントロール
    • ab41927 gave a positive result in Human Lung tissue lysate, Human Brain tissue lysate and Hela whole cell lysate.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

画像

  • 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

参考文献

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 21 Publications for this product

レビューと Q&A

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

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