GFP-Booster (Atto 647N)

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Antigen
  • green fluorescent protein
  • gfp
Reaktivität
Aequorea victoria
Antikörpertyp
Recombinant Antibody
Konjugat
Atto 647N
Applikation
Fluorescence Microscopy (FM), Immunofluorescence (IF)
Optionen
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Verwendungszweck With our Booster you reactivate, boost, stabilizate the signals of your fusion proteins.
Spezifität GFP-Booster efficiently detects and labels most common GFP derivates. No binding to red fluorescent proteins derived from DsRed can be detected.
Produktmerkmale
  • Enhance, stabilize and reactivate your fl uorescent proteins
  • GFP-Booster highly specific for GFP fusion proteins (and derivatives thereof e.g. YFP or Venus)
  • Coupled to bright and photostable chemical dyes from ATTO-TEC
Bestandteile GFP-Trap® coupled to fluorescent dye ATTO 647N
Andere Bezeichnung Green Fluorescent Protein
Hintergrund Green fluorescent proteins (GFP) and variants there of are widely used to study protein localization and dynamics in living cells. However, photo stability and quantum efficiency of GFP 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 GFP signal.The GFP-Booster_Atto488, a specific GFP-binding protein coupled to the fluorescent dye ATTO 594, reactivates, boosts and stabilizes your GFP signal.
Applikationshinweise For the immunofluorescence staining of GFP-fusion proteins in fixed cells
Abberior STAR 635N:
λex 634 nm; λem 654 nm in PBS, pH 7.4, λSTED 750 − 780 nm
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. GFP-Booster incubation: dilute GFP-Booster 1:200 in blocking buffer and incubate 1 h, RT. Note: For multiplexing protocols you can combine GFP-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. Please note: Optimal dilutions/ concentrations should be determined by the end user
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 1 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