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Super Fluor® 488 （～Alexa Fluor 488 ）
Super Fluor 488 Acid
Super Fluor®are registered trademarks and strictly forbidden to be used without authorization！
Super Fluor® 488 (~Alexa Fluor) is a sulfonated rhodamine fluorophore with the same chemical structure of Alexa Fluor 488 for demanding applications including microscopy. Super Flour® 488 is water soluble and pH-insensitive from pH 4 to pH 10. It is a bright yellow-fluorescent dye optimal for use with the 488 nm Argon laser line. The dye emits in green channel, it is compatible with Sybr Green, FAM and FITC filter sets.
| Molecular weight | Excitation (nm) | Emission (nm) | Extinction coefficient (cm-1 M-1) | Quantum yield |
| 648.49 | 499 | 520 | 73000 | 0.92 |
Super Fluor 488，555, 647, 680, 750
|
488 |
555 |
647 |
680 |
750 |
|
|
molecular weight |
～900 |
～1250 |
～1300 |
～1150 |
～1300 |
|
λmax(nm) Ex |
494 |
555 |
647 |
680 |
750 |
|
Molar Absorption Coefficient(M-1cm-1) |
71 000 |
150 000 |
239 000 |
184000 |
240 000 |
|
λmax(nm) Em |
524 |
565 |
665 |
702 |
775 |
|
A280nm /Amax |
11% |
8% |
3% |
5％ |
4% |
Succinimidyl esters are proven to be the best reagents for amine modifications because the amide bonds that are formed are essentially identical to, and as stable as the natural peptide bonds. These reagents are generally stable and show good reactivity and selectivity with aliphatic amines. There are few factors that need be considered when SE compounds are used for conjugation reaction:
1). Solvents: Dissolve NHS ester in 1/10 reaction anhydrous volume of DMF or DMSO.
2). Reaction pH: The labeling reactions of amines with succinimidyl esters are strongly pH dependent. Amine-reactive reagents react with non-protonated aliphatic amine groups, including the terminal amines of proteins and the e-amino groups of lysines. Thus amine acylation reactions are usually carried out above pH 7.5. Protein modifications by succinimidyl esters can typically be done at pH 7.5-8.5, whereas isothiocyanates may require a pH 9.0-10.0 for optimal conjugations.
3). Reaction Buffers: Buffers that contain free amines such as Tris and glycine and thiol compounds must be avoided when using an amine-reactive reagent. Ammonium salts (such as ammonium sulfate and ammonium acetate) that are widely used for protein precipitation must also be removed (such as viadinlysis) before performing dye conjugations.
4). Reaction Temperature: Most conjugations are done at room temperature. However, either elevated or reduced temperature may be required for a particular labeling reaction.
5) Use 10:1 molar ratio Dye NHS/ protein to perform the labeling. Calculate required amount of NHS ester and the protein.
6) Add NHS ester solution to the solution of biomolecule, and vortex well. Keep on ice overnight, or at room temperature during at least 4 hours.
7) Purify the conjugation. The dye-protein conjugate purification by using a Sephadex G-25 column. Add PBS (pH 7.2-7.4) to the desired sample to complete the column purification. Combine the fractions that contain the desired dye-protein conjugate.
8) Determine the optimal dye/protein ratio (optional):Each protein requires distinct dye/protein ratio, which also depends on the properties of dyes. Over labeling of a protein could detrimentally affects its binding affinity while the protein conjugates of low dye/protein ratio gives reduced sensitivity.
Safety warnings and precautions
NOTE: The qualified persons should always wear lab coats, gloves and goggles when working with our products although they are low-risk chemicals for R&D only.
特别提醒：
1) 本公司所有产品仅限于专业人员用于生命科学研究，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅。
2) 本公司所有产品必须由合格专业技术人员操作同时佩戴口罩/手套/实验服并遵守生物实验室安全操作规程！
Description
| Name | Super Fluor® 488 （～Alexa Fluor 488 ） | ||
|---|---|---|---|
| CAT# | 1003-1mg | CAS# | N/A |
| Storage# | −20°C Sealed & desiccated & Minimized light exposure | Shelf Life# | 12个月 |
| Ex(nm)# | 494 | Em(nm)# | 518 |
| MW# | N/A | Solvent# | DMSO |
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