RFP-Booster (Atto 647N)

Details zu Produkt Nr. ABIN2745402, Anbieter: Anmelden zum Anzeigen
Antigen
  • RFP
  • RNF76
  • tripartite motif containing 27
  • TRIM27
Reaktivität
Discosoma
Antikörpertyp
Recombinant Antibody
Konjugat
Atto 647N
Applikation
Fluorescence Microscopy (FM), Immunofluorescence (IF)
Optionen
Hersteller
Anmelden zum Anzeigen
Hersteller Produkt- Nr.
Anmelden zum Anzeigen
Verwendungszweck With our Booster you reactivate, boost, stabilizate the signals of your fusion proteins.
Spezifität RFP-Booster efficiently highlights, enhancesand stabilizes monomeric red fluorescentproteins such as mRFP1, mCherry or mPlum but also mRuby
Produktmerkmale
  • Enhance, stabilize and reactivate your fl uorescent proteins
  • RFP-Booster high specifi city for various monomeric red fl uorescent proteins derived from DsRed
  • Coupled to bright and photostable chemical dyes from ATTO-TEC
Bestandteile RFP-Trap® coupled to fluorescent dye ATTO 647N
Andere Bezeichnung RFP
Hintergrund Red fluorescent proteins (RFP) and variants thereof are widely used to study protein localization and dynamics in living cells. However, photo stability and quantum efficiency of RFP are not sufficient for Super-Resolution Microscopy (e.g. 3D-SIM or STED) of fixed samples. In addition, many cell biological methods such as BrdU-staining, EdU-Click-iT™ treatment or Fluorescent In Situ Hybridization result in disruption of the RFP signal.The RFP-Booster_Atto647N, a specific RFP-binding protein coupled to the fluorescent dye ATTO 647N, reactivates, boosts and stabilizes your RFP signal.
Forschungsgebiet Tags/Labels
Applikationshinweise For the immunofluorescence staining of RFP-fusion proteins in fixed cells
Kommentare

Booster are very small, highly specific GFP- or RFP-binding proteins covalently coupled to the superior fluorescent dyes from ATTO-TEC.

Testdurchführung
  • 1. Fixation: 4% paraformaldehyde (PFA) or 1:10 formalin (37% formaldehyde, 10-15% MetOH) in PBS, 10 min., RT.
  • 2. Wash 3x with PBS containing 0.1% Tween 20 (PBST). Critical: do not let coverslips “dry”.
  • 3. Permeabilisation: PBS containing 0.5% Triton X-100, 5 min., RT. Alternatively, permeabilise by incubating in 100% methanol for 5 min at -20°C.
  • 4. Wash 2x with PBST.
  • 5. Blocking: 4% BSA in PBST, 10 min, RT.
  • 6. RFP-Booster incubation: dilute RFP-Booster 1:200 – 1:400 in blocking buffer and incubate 1 h, RT.
    Note: For multiplexing protocols you can combine RFP-Booster with any other antibody.
  • 7. Wash 3x 5-10 min in PBST.
  • 8. If required, counterstain with DNA fluorescent dyes, e.g. DAPI.
  • 9. Before mounting, coverslips can be very briefly rinsed in water to prevent salt crystals to form.
  • 10. Mount in VectaShield (Vector Labs) or other mounting media with anti-fading agents and seal mounted coverslips with clear nail polish.
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 0.2 mg/ml
Buffer PBS, 0.01% Sodium azide
Konservierungsmittel Sodium azide
Vorsichtsmaßnahmen This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung Do not freeze. Protect from light.
Lagerung 4 °C
Haltbarkeit 6 months