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GFP-Booster (Atto 488)

Details zu Produkt Nr. ABIN509419, Anbieter: Anmelden zum Anzeigen
Antigen
Reaktivität
Aequorea victoria
Antikörpertyp
Recombinant Antibody
Konjugat
Atto 488
Applikation
Fluorescence Microscopy (FM), Immunofluorescence (IF)
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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 specifi c 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 488
Andere Bezeichnung GFP
Hintergrund 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.
Forschungsgebiet Tags/Labels
Applikationshinweise For the immunofluorescence staining of GFP-fusion proteins in fixed cells

ATTO 488:
Excitation range 480 - 510 nm (λabs= 501 nm)
Emission range 520 - 560 nm (λfl= 523 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 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
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
Bilder des Herstellers
 image for GFP-Booster (Atto 488) (ABIN509419) Enhancement of GFP signal with GFP-Booster after EdU-Click-iT™ treatment. EdU-Click-i...
Immunofluorescence (IF) image for GFP-Booster (Atto 488) (ABIN509419) Visualization of GFP signal with GFP-Booster after EdU-Click-iT™ treatment. EdU-Click...
 image for GFP-Booster (Atto 488) (ABIN509419) Enhancement of GFP signal with GFP-Booster_Atto488. Comparison of signal intensity of...
Produkt verwendet in: Vleugel, Omerzu, Groenewold, Hadders, Lens, Kops: "Sequential multisite phospho-regulation of KNL1-BUB3 interfaces at mitotic kinetochores." in: Molecular cell, Vol. 57, Issue 5, pp. 824-35, 2015

Eikenes, Malerød, Christensen, Steen, Mathieu, Nezis, Liestøl, Huynh, Stenmark, Haglund: "ALIX and ESCRT-III coordinately control cytokinetic abscission during germline stem cell division in vivo." in: PLoS genetics, Vol. 11, Issue 1, pp. e1004904, 2015

Vietri, Schink, Campsteijn, Wegner, Schultz, Christ, Thoresen, Brech, Raiborg, Stenmark: "Spastin and ESCRT-III coordinate mitotic spindle disassembly and nuclear envelope sealing." in: Nature, Vol. 522, Issue 7555, pp. 231-5, 2015

Qin, Wolf, Liu, Link, Smets, Mastra, Forné, Pichler, Hörl, Fellinger, Spada, Bonapace, Imhof, Harz, Leonhardt: "DNA methylation requires a DNMT1 ubiquitin interacting motif (UIM) and histone ubiquitination." in: Cell research, Vol. 25, Issue 8, pp. 911-29, 2015

Vleugel, Hoek, Tromer, Sliedrecht, Groenewold, Omerzu, Kops: "Dissecting the roles of human BUB1 in the spindle assembly checkpoint." in: Journal of cell science, Vol. 128, Issue 16, pp. 2975-82, 2015

Riemer, Bontems, Krishnakumar, Gömann, Dosch: "A functional Bucky ball-GFP transgene visualizes germ plasm in living zebrafish." in: Gene expression patterns : GEP, Vol. 18, Issue 1-2, pp. 44-52, 2015

Seybold, Elserafy, Rüthnick, Ozboyaci, Neuner, Flottmann, Heilemann, Wade, Schiebel: "Kar1 binding to Sfi1 C-terminal regions anchors the SPB bridge to the nuclear envelope." in: The Journal of cell biology, Vol. 209, Issue 6, pp. 843-61, 2015

Richens, Barros, Lucas, Peel, Pinto, Wainman, Raff: "The Drosophila Pericentrin-like-protein (PLP) cooperates with Cnn to maintain the integrity of the outer PCM." in: Biology open, Vol. 4, Issue 8, pp. 1052-61, 2015

Hall, Keighren, Ford, Davey, Jarman, Smith, Jackson, Mill: "Acute versus chronic loss of mammalian Azi1/Cep131 results in distinct ciliary phenotypes." in: PLoS genetics, Vol. 9, Issue 12, pp. e1003928, 2014

Vleugel, Tromer, Omerzu, Groenewold, Nijenhuis, Snel, Kops: "Arrayed BUB recruitment modules in the kinetochore scaffold KNL1 promote accurate chromosome segregation." in: The Journal of cell biology, Vol. 203, Issue 6, pp. 943-55, 2014

Winterhoff, Junemann, Nordholz, Linkner, Schleicher, Faix: "The Diaphanous-related formin dDia1 is required for highly directional phototaxis and formation of properly sized fruiting bodies in Dictyostelium." in: European journal of cell biology, Vol. 93, Issue 5-6, pp. 212-24, 2014

Agircan, Schiebel: "Sensors at centrosomes reveal determinants of local separase activity." in: PLoS genetics, Vol. 10, Issue 10, pp. e1004672, 2014

Biermann, Sokoll, Klueva, Missler, Wiegert, Sibarita, Heine: "Imaging of molecular surface dynamics in brain slices using single-particle tracking." in: Nature communications, Vol. 5, pp. 3024, 2014

Bleck, Itano, Johnson, Thomas, North, Bieniasz, Simon: "Temporal and spatial organization of ESCRT protein recruitment during HIV-1 budding." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 111, Issue 33, pp. 12211-6, 2014

Bluteau, Zhuang, Amann, Trueb: "Targeted disruption of the intracellular domain of receptor FgfrL1 in mice." in: PLoS ONE, Vol. 9, Issue 8, pp. e105210, 2014

Bourge, Fort, Soler, Satiat-Jeunemaître, Brown: "A pulse-chase strategy combining click-EdU and photoconvertible fluorescent reporter: tracking Golgi protein dynamics during the cell cycle." in: The New phytologist, 2014

Britton, Dernoncourt, Delteil, Froment, Schiltz, Salles, Frit, Calsou: "DNA damage triggers SAF-A and RNA biogenesis factors exclusion from chromatin coupled to R-loops removal." in: Nucleic acids research, Vol. 42, Issue 14, pp. 9047-62, 2014

Kruse, Larsen, Sedgwick, Sigurdsson, Streicher, Olsen, Nilsson: "A direct role of Mad1 in the spindle assembly checkpoint beyond Mad2 kinetochore recruitment." in: EMBO reports, Vol. 15, Issue 3, pp. 282-90, 2014

Linkner, Witte, Zhao, Junemann, Nordholz, Runge-Wollmann, Lappalainen, Faix: "The inverse BAR domain protein IBARa drives membrane remodeling to control osmoregulation, phagocytosis and cytokinesis." in: Journal of cell science, Vol. 127, Issue Pt 6, pp. 1279-92, 2014

Oliveira, Kotadia, Tavares, Mirkovic, Bowlin, Eichinger, Nasmyth, Sullivan: "Centromere-Independent Accumulation of Cohesin at Ectopic Heterochromatin Sites Induces Chromosome Stretching during Anaphase." in: PLoS biology, Vol. 12, Issue 10, pp. e1001962, 2014