BHQ-3 Carboxylic Acid
BHQ-3 Carboxylic Acid
Key featuresShow Hide
- Used to conjugate a non-fluorescent quencher to an amine functionality of a biomolecule.
- Maximal absorption in the 620 to 730 nm range.
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Since their introduction to the DNA marketplace in 2000, the Black Hole Quenchers have become THE gold-standard quencher of choice used in qPCR probes and other FRET applications. When fluorogenic dual-labeled probes were first introduced, quenchers were often a second reporter dye, typically TAMRA. The sensitivity of these probes, such as FAM-TAMRA, is limited because the fluorescence from TAMRA can leak into the channel meant to detect FAM fluorescence. Dark quenchers, which are dyes with no native fluorescence, offer a solution to this problem. The BHQ dyes are true dark quenchers with no native emission due to their polyaromatic-azo backbone. Substituting electron-donating and withdrawing groups on the aromatic rings produces a complete series of quenchers with broad absorption curves that span the visible spectrum into the near IR region. BHQ dyes work through a combination of FRET and static quenching to enable researchers to avoid the residual background signal common to fluorescing quenchers such as TAMRA, or low signal to noise ratio. These quenchers can be paired with all common reporter dyes to construct efficiently quenched qPCR probes for multiplexing assays. In addition to quenching by FRET, BHQ dyes have also been shown to efficiently quench fluorescence through static quenching via formation of a ground state complex with the reporter dye. BHQ quenchers have broad absorption spectra and can be paired with reporter dyes that emit in the following ranges: BHQ-0: 430-520 nm BHQ-1: 480-580 nm BHQ-2: 560-670 nm BHQ-3: 620-730 nm BHQ-10: 480-550 nm Water Soluble (WS) LGC, Biosearch Technologies offers all Black Hole Quencher products available for 3’, internal and 5’ modification of oligonucleotides with a variety of options. BHQ-1 and BHQ-2 are the more popular, either as the 5'-Phosphoramidites, the dT-Phosphoramidites or the 3'-CPGs. Only considering the excitation and emission values suggests Cy™5/Cyanine-5 and Quasar 670 require BHQ-3 for efficient quenching, however BHQ-2 is recommended because it is less susceptible to degradation. BHQ-1 is typically used to quench in the range 480 - 580 nm and can be used in conjunction with the commonly used fluorophores; e.g. FAM, TET, JOE and HEX. BHQ-2 is used to quench in the range 550 – 650 nm and is most effective in quenching fluorophores such as TAMRA, ROX, Cyanine-3, Cy3, Cy3.5™ and Red 640. We also offer Black Hole Quenchers for labelling peptides. All of our BHQ dyes are available as carboxylic acid or succinimidyl esters. Our BHQ-1 and BHQ-2 dyes are available as FMOC lysine conjugates. Our water soluble BHQ-10 is available as a carboxylic acid or succinimidyl ester.
- Formula: C33H35F6N6O2P
- Molecular Weight: 692.63
- Appearance: dark purple solid
- Absorption Maximum (Lambda Max): 615
- Extinction Coefficient at Lambda max: 40700
- Extinction Coefficient at 260 nm: 13000
Spectral properties measured in phosphate-buffered saline (PBS) pH 7.3.
- Product Solubility:This product can be dissolved in methanol, DMSO, MeCN or DMF.Additional Spectroscopic Data:The absorption maximum of BHQ-3 is dependent on environmental factors such as solvent and the nature of the substrate to which it is attached.Absorption max. in methanol: 656 nmAbsorption max. in DCM: 678 nmExtinction Coefficient at 280 nm (PBS, pH 7.3): 25,000/M/cmFor BHQ-3 dye conjugated to the 5' end of a poly T oligonucleotide:Extinction Coefficient at absorption max, 670 nm, (PCR buffer): 42,700/M/cmProduct Stability:Reducing agents that are commonly used to reduce dithiol bonds in proteins, such as DTT and TCEP, will also reduce BHQ-3 thereby causing a loss of color and of quenching efficiency.
Storage and handling:
- Shipping conditions: Ambient
- Storage conditions: -15 to -30 °C