- Antibody Type
- Recombinant Antibody
- Aequorea victoria
- Atto 488
- Fluorescence Microscopy (FM), Immunofluorescence (IF)
- With our Booster you reactivate, boost, stabilizate the signals of your fusion proteins.
- GFP-Booster efficiently detects and labels most common GFP derivates. No binding to red fluorescent proteins derived from DsRed can be detected.
- Enhance, stabilize and reactivate your fl uorescent proteins
- GFP-Booster highly specifi c for GFP fusion proteins (and derivatives thereof e.g. YFP or Venus)
- Coupled to bright and photostable chemical dyes from ATTO-TEC
- GFP-Trap® coupled to fluorescent dye ATTO 488
- Application Notes
For the immunofluorescence staining of GFP-fusion proteins in fixed cells
Excitation range 480 - 510 nm (λabs= 501 nm)
Emission range 520 - 560 nm (λfl= 523 nm)
Booster are very small, highly specific GFP- or RFP-binding proteins covalently coupled to the superior fluorescent dyes from ATTO-TEC.
- Assay Procedure
- 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 5min 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
- For Research Use only
- 1 mg/ml
- PBS, 0.01% Sodium azide
- Sodium azide
- Precaution of Use
- This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Handling Advice
- Do not freeze. Protect from light.
- 4 °C
- Expiry Date
- 6 months
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- Rehage, Davydova, Conrad, Behrens, Maiser, Stehklein, Brenner, Klein, Jeridi, Hoffmann, Lee, Dianzani, Willemsen, Feederle, Reiche, Hackermüller, Leonhardt, Sharma, Niessing, Heissmeyer: "Binding of NUFIP2 to Roquin promotes recognition and regulation of ICOS mRNA." in: Nature communications, Vol. 9, Issue 1, pp. 299, 2018 (PubMed).
- Alternative Name
- green fluorescent protein, gfp
- Green fluorescent proteins (GFP) and variants thereof 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 488, reactivates, boosts and stabilizes your GFP signal.