sgGFP™: SuperGlo™ Green Fluorescent Protein

• Broadest Stoke's shift of all green variants
• Single enhanced excitation peak

Mutations present in sgGFP™:

• F64L: improved solubility, and protein folding at 37°C;
  more rapid and efficient fluorophore formation
• S65C: a single excitation peak at 474 nm, and emission at 509 nm
• I167T: enhanced excitation peak at 474 nm

Qbiogene's sgGFP™ is the only available GFP with a single
enhanced excitation peak at 474 nm and a green fluorescence at ~ 509 nm. All other popular variants have excitation peaks at or near 488 nm. sgGFP™ is the only GFP that has a Stoke's shift that exceeds the minimum of 30 nm recommended for easy visualization and sensitive detection. sgGFP™ can be readily detected in fluorimeters, fluorescent cell sorters (FACS) and by fluorescent microscopes. sgGFP™ is also the best partner for FRET with sgBFP™.

sgBFP™: SuperGlo™ Blue Fluorescent Protein

• Highest quantum yield of blue variants.

Mutations present in sgBFP™:

• F64L: improved solubility, and protein folding at 37°C;
  more rapid and efficient fluorophore formation
• Y66H: a single excitation peak at 387 nm, and emission at 450 nm
• V163A: improved solubility and protein folding

Qbiogene's sgBFP™ is the perfect partner for sgGFP™. The Y66H mutation generates the spectral shift to a blue emitting variant. The unique combination of the F64L, Y66H and V163A mutations present in sgBFP™ leads to a QY (Quantum Yield) that approaches the QY of the brightest green variants and is 2- 3 fold higher than the QY of mutants without S65T. sgBFP™ is also more resistant to photobleaching than other BFPs and its spectral properties make it easy to distinguish from GFP or YFP. sgBFP™ can be readily detected in fluorimeters, FACs and by fluorescent microscopes. Furthermore, the large overlap between the emission peak of sgBFP™ and the excitation peak of sgGFP™ make them perfect partners for FRET.

Purified Recombinant AFP^® Proteins
Purified recombinant SuperGlo™ GFP and SuperGlo™ BFP proteins are available. These can be used as standards to calculate expression levels and correlated to relative fluorescence intensities.

storage: -20° C

Antibodies for detecting GFP, BFP and other variants of wtGFP

Two monoclonal antibodies (mAb) are offered for the detection of Qbiogene’s SuperGlo™ GFP and SuperGlo™ BFP. Both recognize also all other variants. Anti-AFP^® mAbs are purified from the supernatant of mouse hybridomas.

The 11E5 mAb, which is a mouse IgG1 monoclonal antibody recognizing GFP and other variants in their denatured form, is ideal for detection in Western analyses. Use of the 11E5 mAb allows detection of as little as 10 ng of GFP using colorimetric methods, and will detect picogram quantities with chemiluminescent
detection assays. Performing Western blots with the
11E5 mAb allows confirmation of AFP^® or AFP^®-fusion protein expression at the expected molecular weight.

The 3E6 mAb, an IgG2a antibody recognizing the native form of GFP, can be used for immuno-precipitation, immunocytochemistry, and ELISA assays. Purification of AFP^® and AFP^®- fusion proteins is possible using either protein A or protein G.

Both antibodies are highly specific for GFP and its variants in both bacterial and mammalian cell extracts, giving strong, clean signals.

storage: -80° C