Chromotek-RFP-Trap Booster

Website: www.allelebiotech.com Call: 1-800-991-RNAi/858-587-6645 (Pacic Time: 9:00AM~5:00PM) Email: oligo@allelebiotech.com

Description Specicity

RFP-Trap coupled to uorescent dye ATTO 594 RFP-Trap efciently highlights monomeric derivates of dsRed (mRFP1, mCherry,mOrange, mPlum), but not GFP and its variants. Red fluorescent proteins (RFPs) are widely used as fluorescent tags. Commonly, a combination of red and green fusion proteins (GFPs) is employed to allow multicolor tracking and analysis of protein-protein interactions. In most applications, the ability to observe fluorescence from red florescent proteins is hindered because the quantum yield and photostability of RFPs are usually not sufficient enough for their microscopic detection. This limitation is particularly important for Super Resolution Microscopy applications (e.g. 3D-SIM or STED). Many cell biological methods such as HCl treatment for BrdU-detection, the EdU-Click-iT treatment or heat denaturation for FISH, lead to disruption of the RFP signal. By using RFP-Booster_Atto594, a specific RFP-binding protein coupled to the fluorescent dye ATTO 594 (from ATTO-TEC), your RFP signal will significantly increase in stability and brightness.
ATTO 594 is a fluorescent label belonging to the class of Rhodamine dyes. Fluorescence is excited most efficiently in the range 560 - 615 nm (abs= 601 nm), a standard 561 nm laser is also suitable for excitation. Maximum fluorescence emission falls in the range 615 - 680 nm (fl= 627 nm).

Relevance

For Technical Support: Email:FP@allelebiotech.com

or Research Use Only. Not for Diagnostic or Therapeutic Use. Purchase does not include or carry any right to resell or transfer this product either as a stand-alone product or as a component of another product. Any use of this product other than the permitted use without the express written authorization of Allele Biotech is strictly prohibited

For further information please refer to http://www.atto-tec.com

Tested Applications

Immunouorescence

HeLa cells expressing mRFP-PCNA fixed and permeabilized. Staining with RFPBooster_Atto594 (dilution 1/200, 1 h at RT).

Conc. Storage

1 mg/ml Shipped at RT. Store at 4C Avoid freezing

Product List Chromotek-RFP-Trap Booster ACT-CM-RBOOSTR 100 ug

Protocols
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 -20C. 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.* 7. Wash 3x 5-10 min in PBST. 8. If required counterstain with DNA uorescent dyes, e.g. DAPI. 9. Before mounting, briey rince coverslipsbe very rinsed in water to prevent salt crystal formation. 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. Tip: RFP-Booster can be added to master-mix during routine immunostaining, along with the primary or secondary antibodies.

Allele Biotech-Introducing Cost Effectiveness to Research