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Flotillin 1 anticorps (C-Term)

FLOT1 Reactivité: Humain, Boeuf (Vache), Chien, Porc WB, IHC (p) Hôte: Chèvre Polyclonal unconjugated
N° du produit ABIN374222
  • Antigène Voir toutes Flotillin 1 (FLOT1) Anticorps
    Flotillin 1 (FLOT1)
    Épitope
    • 8
    • 7
    • 6
    • 4
    • 4
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    C-Term
    Reactivité
    • 60
    • 28
    • 27
    • 7
    • 6
    • 6
    • 5
    • 4
    • 4
    • 3
    • 3
    • 2
    • 1
    • 1
    • 1
    Humain, Boeuf (Vache), Chien, Porc
    Hôte
    • 56
    • 3
    • 2
    Chèvre
    Clonalité
    • 48
    • 13
    Polyclonal
    Conjugué
    • 30
    • 7
    • 4
    • 4
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    Cet anticorp Flotillin 1 est non-conjugé
    Application
    • 39
    • 23
    • 10
    • 7
    • 6
    • 3
    • 2
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
    Specificité
    This antibody detetcs (FLOT1) at C-term.
    Réactivité croisée (Details)
    Species reactivity (expected):Dog, Pig, Cow.
    Species reactivity (tested):Human.
    Purification
    Affinity chromatography
    Immunogène
    ptide with sequence C-SISQVNHKPLRTA, from the C Terminus of the protein sequence Genename: FLOT1
    Top Product
    Discover our top product FLOT1 Anticorps primaire
  • Indications d'application
    Peptide ELISA: 1/32000. Western blot: 0,3 - 1,0 1 μg/mL. Approx 48 kDa band observed in H460 lysates (predictedMW of 48 kDa according to NP_005794).
    Other applications not tested.
    Optimal dilutions are dependent on conditions and should be determined by the user.
    Restrictions
    For Research Use only
  • Validation #101253 (Western Blotting)
    'Independent Validation' signe
    by
    Internal Disease Clinic, Faculty of Veterinary Medicine, University of Zagreb
    No.
    #101253
    Date
    03.05.2017
    Antigène
    Flotillin-1
    Numéro du lot
    Application validée
    Western Blotting
    Contrôle positif
    extracellular vesicles from bovine follicular fluid, verified with CD63 antibody
    Contrôle négative
    non-enriched fractions
    Conclusion
    Passed. The flotillin-1 antibody ABIN374222 specifically reveals a band of the expected molecular weight in fractions enriched for exosomes.
    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    ABIN374222
    Anticorps secondaire
    anti-goat-HRP
    Full Protocol
    • Isolate extracellular vesicles from 0.5ml follicular fluid from cow obtained from abattoir using the qEV size-exclusion chromatography kit (iZON Science, qEVoriginal Size Exclusion Columns, NC0888135):
      • Equilibrate columns with 10ml PBS.
      • Apply the follicular fluid.
      • Retain the first 3ml of the eluate as void volume negative control (V0).
      • Collect the next three 0.5ml fractions (1, 2, 3) which are expected to contain extracellular vesicles.
      • Collect next 0.5ml fraction (4) which is expected to contain proteins but not extracellular vesicles from follicular fluid.
    • Concentrate fractions 25x using Microcon filters (Millipore, Ultracel YM-30, MRCF0R30) with a 25kDa cut-off.
    • Boil all five fractions and 1µl of untreated follicular fluid as positive control in 1x Laemmli SDS sample buffer for 5min.
    • Separate all protein in each fraction, 10µg for fraction 2, the richest in protein on a freshly cast 4% acrylamide stacking gel and 10% acrylamide separation SDS-PAGE gel.
    • Transfer proteins onto a nitrocellulose membrane (GE Healthcare,Amersham Protran 0.45µm, 10600002) in an electroblotter tank (Biostep, electroblotting module GV100- EBGRM) Western blotting system using 20% methanol-containing transfer buffer for 2h at 150mA at RT.
    • Block the membrane with Odyssey blocking buffer (PBS) (Odyssey, 927-400000) for 1h at RT.
    • Incubation with primary goat anti-flotillin 1 antibody (antibodies-online, ABIN374222) diluted 1:500 in blocking buffer ON at 4°C.
    • Wash membrane 3x 5min with TBST (TBS, 0.1% Tween-20).
    • Incubate with secondary anti-goat-HRP antibody diluted 1:5000 in blocking buffer for 1h at RT.
    • Wash membrane 3x 5min with TBST.
    • Incubate membrane in Luminol for (Santa Cruz Biotechnology Inc., ImmunoCruz western blotting luminol reagent, SC-2048) 5min at RT.
    • Reveal chimiuminescent bands using a chemiluminescence detector (LI-COR, Odyssey Fc, OFC-0966) imaging system.
    • Strip membranes usingmild stripping protocol:
      • Incubate membrane with stripping buffer (1.5% (w/v) glycine, 0.0% (w/v) SDS, 1% (v/v) tween 20, pH2.2) shaking for 2x 10min at RT.
      • Wash membrane 2x 10min with PBS.
      • Wash membrane2x 5min in TBS.
      • Block the membrane with blocking buffer.
    • Incubate with primary goat anti-CD63 antibody (antibodies-online, ABIN1440014, lot 0047080912) diluted 1:200 in blocking buffer ON at 4°C.
    • Repeat the same steps as for the goat anti-flotillin 1 antibody.
    Notes
    We observed an albumin band in western blot of some fractions when we used a different secondary (HRP conjugated mouse anti-CD9 antibody) (not shown). Albumin was not observed with ABIN374222.
  • Concentration
    0.5 mg/mL
    Buffer
    Tris saline, pH ~7.3, 0.02 % Sodium Azide, 0.5 % BSA
    Agent conservateur
    Sodium azide
    Précaution d'utilisation
    This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Conseil sur la manipulation
    Avoid repeated freezing and thawing.
    Stock
    4 °C/-20 °C
    Stockage commentaire
    Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
  • Antigène
    Flotillin 1 (FLOT1)
    Autre désignation
    Flotillin-1 / FLOT1 (FLOT1 Produits)
    Synonymes
    anticorps reggie-2, anticorps flot1, anticorps Rareg, anticorps flot1c, anticorps flotillin-1, anticorps flotillin1, anticorps flot1a, anticorps reggie2, anticorps si:dz44o19.1, anticorps wu:fj45g11, anticorps flotillin 1, anticorps flotillin-1, anticorps putative flotillin-1, anticorps flotillin 1 S homeolog, anticorps flotillin 1 L homeolog, anticorps flotillin 1a, anticorps FLOT1, anticorps Flot1, anticorps flot1, anticorps LOC5566722, anticorps CpipJ_CPIJ008623, anticorps Smp_016200.1, anticorps flot1.S, anticorps flot1.L, anticorps flot1a
    Sujet
    Flotillin-1 is a lipid raft-associated protein that has been implicated in various cellular processes: it has been reported to function as a molecular link between lipid rafts of the plasma membrane and a multimeric signaling complex at the actin cytoskeleton, to associate with caveolae , regulating vesicular trafficking and signal transduction, and to have mitogenic properties. Its subcellular localisation seems to be cell-type-specific. It has been found associated with the cell membranes, nucleus, cytoplasm and different cytoplasmic organelles.Synonyms: REG-2, Reg2, Reggie-2
    ID gène
    10211
    NCBI Accession
    NP_005794
    UniProt
    O75955
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