GFP-Booster_atto488-TS

Application: Immunofluorescence (IF), Immunhistochemie (IHC)

Catalog no.: ABIN1082212

Additional Information

Description: GFP-Trap® coupled to fluorescent dye ATTO 488

Specificity: GFP-Booster efficiently detects and labels most common GFP derivates. No binding to red fluorescent proteins derived from DsRed can be detected.

Comments:

Booster are very small, highly specific GFP- or RFP-binding proteins covalently coupled to the superior fluorescent dyes from ATTO-TEC.
With our Booster you reactivate, boost, stabilizate the signals of your fusion proteins.

Application Details

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

Application Notes: GFP-Booster now enables you to use your existing GFP expression constructs and cell lines also for super-resolution microscopy.

Concentration: 1 mg/ml

Buffer: PBS

Preservative: 0.01% Sodium azide

Storage: Shipped at ambient temperature. Upon receipt store at 4°C. Do not freeze. Protect from light.

Restrictions: For Research Use only

Images

Visualization of GFP signal with GFP-Booster after EdU-Click-iT™ treatment.EdU-Click-iT™ treament leads to disruption of GFP signal. GFP-Booster labels GFP fusion proteins and thus reactivates and boosts the fluorescence (dilution 1/200,1 h at RT).(GFP-Booster with EdU-Click-iT™ protocol)

Enhancement of GFP signal with GFP-Booster_Atto488. Comparison of signal intensity of a HeLa cell line stably expressing a nuclear GFP-fusion protein before and after GFP-Booster treatment.

Publications

Product: Guizetti, Schermelleh, Mäntler et al.: "Cortical constriction during abscission involves helices of ESCRT-III-dependent filaments." in: Science (New York, N.Y.), Vol. 331, Issue 6024, pp. 1616-20, 2011 (PubMed).

Cordes, Maiser, Steinhauer et al.: "Mechanisms and advancement of antifading agents for fluorescence microscopy and single-molecule spectroscopy." in: Physical chemistry chemical physics : PCCP, Vol. 13, Issue 14, pp. 6699-709, 2011 (PubMed).

Ridzuan, Moon, Knuepfer et al.: "Subcellular location, phosphorylation and assembly into the motor complex of GAP45 during Plasmodium falciparum schizont development." in: PLoS ONE, Vol. 7, Issue 3, pp. e33845, 2012 (PubMed).

Mikeladze-Dvali, von Tobel, Strnad et al.: "Analysis of centriole elimination during C. elegans oogenesis." in: Development (Cambridge, England), Vol. 139, Issue 9, pp. 1670-9, 2012 (PubMed).

Ries, Kaplan, Platonova et al.: "A simple, versatile method for GFP-based super-resolution microscopy via nanobodies." in: Nature methods, Vol. 9, Issue 6, pp. 582-4, 2012 (PubMed).