RFP-Booster (Atto 647N)

ABIN2745402 产品详细信息, 供应商: Log in to see
抗原
适用
Discosoma
抗体类型
Recombinant Antibody
标记
Atto 647N
应用范围
Fluorescence Microscopy (FM), Immunofluorescence (IF)
选项
Supplier
Log in to see
Supplier Product No.
Log in to see
原理 With our Booster you reactivate, boost, stabilizate the signals of your fusion proteins.
特异性 RFP-Booster efficiently highlights, enhancesand stabilizes monomeric red fluorescentproteins such as mRFP1, mCherry or mPlum but also mRuby
产品特性
  • 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
组件 RFP-Trap® coupled to fluorescent dye ATTO 647N
别名 RFP
背景 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_Atto647N, a specific RFP-binding protein coupled to the fluorescent dye ATTO 647N, reactivates, boosts and stabilizes your RFP signal.
研究领域 Tags/Labels
应用备注 For the immunofluorescence staining of RFP-fusion proteins in fixed cells
说明

Booster are very small, highly specific GFP- or RFP-binding proteins covalently coupled to the superior fluorescent dyes from ATTO-TEC.

实验流程
  • 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.
限制 仅限研究用
状态 Liquid
浓度 0.2 mg/ml
缓冲液 PBS, 0.01% Sodium azide
储存液 Sodium azide
注意事项 This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
注意事项 Do not freeze. Protect from light.
储存条件 4 °C
有效期 6 months