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RFP-Booster (Atto 594)

Details for Product No. ABIN1082215
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Target Name (Antigen)
Reactivity
Discosoma
Conjugate
Atto 594
Application
Immunofluorescence (IF), Fluorescence Microscopy (FM)
Pubmed 6 references available
Quantity 50 µg
Options
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Availability Will be delivered in 4 to 5 Business Days
Catalog No. ABIN1082215
178.20 $
Plus shipping costs $45.00

Order hotline:

  • +1 404 474 4654
  • +1 888 205 9894 (TF)
Purpose With our Booster you reactivate, boost, stabilizate the signals of your fusion proteins.
Specificity RFP-Booster efficiently highlights, enhancesand stabilizes monomeric red fluorescentproteins such as mRFP1, mCherry or mPlum but also mRuby
Characteristics - 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
Components RFP-Trap® coupled to fluorescent dye ATTO 594
Alternative Name RFP
Background 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_Atto594, a specific RFP-binding protein coupled to the fluorescent dye ATTO 594, reactivates, boosts and stabilizes your RFP signal.
Research Area Tags/Labels
Application Notes For the immunofluorescence staining of RFP-fusion proteins in fixed cells

ATTO 594:
Excitation range 560 - 615 nm (λabs = 601 nm)
Emission range 615 - 680 nm (λfl= 627 nm)
Comment

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 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.
Restrictions For Research Use only
Concentration 0.2 mg/ml
Buffer PBS, 0.01% Sodium azide
Preservative 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.
Storage 4 °C
Expiry Date 6 months
Supplier Images
RFP-Booster (Atto 594) RFP-Booster specifically labels RFP-fusion proteins. HeLa cells expressing mRFP-PCNA, fixed and permeabilized. Specific staining of early, mid and late replication foci with RFP-Booster_Atto594 (dilution 1/200, 1 h at RT).
RFP-Booster (Atto 594) (2) Enhancement of RFP signal with RFP-Booster_Atto594. Comparison of signal intensity of a HeLa cell line stably expressing a nuclear RFP-fusion protein before and after RFP-Booster treatment (1/200 dilution).
RFP-Booster (Atto 594) (3) Improvement of RFP signal stability with RFP-Booster_Atto594. RFP fluorescence bleaches very quickly upon irradiation with high laser intensity. In contrast, fluorescent signal remains stable when enhanced with RFP-Booster_Atto594.
Product cited in: 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).

Hasegawa, Ryu, Kaláb: "Chromosomal gain promotes formation of a steep RanGTP gradient that drives mitosis in aneuploid cells." in: The Journal of cell biology, Vol. 200, Issue 2, pp. 151-61, 2013 (PubMed).

Request Want additional data for this product?

The Independent Validation Initiative strives to provide you with high quality data. Find out more

Catalog No. ABIN1082215
178.20 $
Plus shipping costs $45.00

Order hotline:

  • +1 404 474 4654
  • +1 888 205 9894 (TF)
Validation Images
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