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AccuOrange™ Protein Quantitation Kit

Highly sensitive, fluorescence-based quantitation of purified proteins.

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Trial size (200 assays)
2,000 assays
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Product Description

AccuOrange™ Protein Quantitation Kit is a highly sensitive fluorescence-based assay for quantifying purified protein samples in 96-well format.

  • Highly sensitive: detect 0.1-15 ug/mL protein
  • Excellent linearity and low variability
  • Stable fluorescence signal
  • Compatible with reducing agents and other additives
  • For use with purified protein or antibody samples
  • Ex/Em: 480/598 nm

The detection range of the assay is 0.1-15 ug/mL protein. AccuOrange™ is much more sensitive than traditional protein quantitation assays such as BCA, Bradford and Lowry, and shows superior linearity and reproducibility compared to the NanoOrange® protein quantitation assay. The assay shows minimal variability between different proteins, and has stable fluorescence signal for up to 16 hours.

AccuOrange™ is recommended for quantifying purified protein or antibody samples. The tolerance of the AccuOrange™ assay to salts, buffers, detergents, and other chemicals is listed in the product protocol. The AccuOrange™ assay has low tolerance for non-ionic detergents, and is not recommended for use with cell lysates containing Triton™ X-100, sodium deoxycholate, CHAPs, or other non-ionic detergents. The assay can tolerate up to 0.01% SDS (final concentration in assay).

Comparison of AccuOrange™ with other protein quantitation assays

AssayDetection RangeComments
AccuOrange™0.1-15 ug/mLFluorescence-based (480/598 nm)
Highly linear
Signal stable for at least 16 hours
Compatible with reducing agents
Not compatible with detergents
NanoOrange®0.1-10 ug/mLFluorescence-based (470/570 nm)
Non-linear
Fluorescence stable for 6 hours
Compatible with reducing agents
Not compatible with detergents
Modified Lowry1-1500 ug/mLAbsorbance-based (750 nm)
Non-linear
Not compatible with reducing agents or detergents
BCA20-2000 ug/mLAbsorbance-based (562 nm)
Highly linear
Signal not stable over time
Not compatible with reducing agents
Compatible with detergents
Bradford (Coomassie®)50-500 ug/mLAbsorbance-based (595 nm)
Signal not stable over time
Non-linear
Compatible with reducing agents
Not compatible with detergents
Pierce® 660 nm50-2000 ug/mLAbsorbance-based (660 nm)
Non-linear
Compatible with reducing agents & detergents
A28050-2000 ug/mLAbsorbance-based (280 nm)
High protein-protein variability
Contaminants and detergents can affect results
NanoOrange is a registered trademark of Thermo Fisher Scientific. Triton is a trademark of The Dow Chemical Company.

Product Attributes

Size
Trial size (200 assays), 2,000 assays
Excitation/Emission
480/598 nm
Storage Conditions
Store at room temperature, Do not refrigerate, Protect dye component from light

Documents, Protocols, SDS and COA

Citations

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FAQs

Protein Detection & Analysis

Even though AccuOrange™ buffer does contain SDS, which is required for the dye to bind proteins, the assay is very sensitive to small changes in SDS concentration, and also cannot tolerate non-ionic detergents that form mixed micelles with SDS, like Triton®. Therefore we don't recommend using the kit for cell lysates or other samples with significant amounts of detergents.

Gels stained with One-Step Blue® can be dried just like gels stained with Coomassie. The stain will not interfere with the detection of radiolabeled proteins.

The AccuOrange™ assay is a fluorescent dye-based assay. The dye binds to proteins primarily through hydrophobic interactions. Proteins denature upon heating; the dye binds to the exposed hydrophobic pockets of the protein after cooling. The free AccuOrange™ dye is fluorogenic due to non-radioactive decay but becomes highly fluorescent due to the rigid conformation inside the pocket.

The AccuOrange™ assay more sensitive than traditional protein quantitation assays such as BCA, Bradford and Lowry, and shows superior linearity and reproducibility than the NanoOrange® protein quantitation assay (Thermo Fisher Sci.), but has low tolerance for detergents like SDS and Triton® X-100.

 

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