pHAb Amine Reactive Dye

pHAb Dyes are pH sensor dyes that have very low fluorescence at pH > 7 and a dramatic increase in fluorescence as the pH of the solution becomes acidic. pHAb Dyes have excitation maxima (Ex) at 532nm and emission maxima (Em) at 560nm.

pHAb Reactive Dyes are designed specifically for antibody labeling and can be used for high-throughput antibody internalization screening assays.

pHAb amine-reactive dyes contain an N-hydroxysuccinimide (NHS) ester group, which reacts with primary amine groups present on lysine residues of antibodies.

A key feature of pHAb dyes is that each dye molecule contains two sulfonate groups. This enhances their solubility in aqueous solutions and reduces aggregation, a common issue observed with non-sulfonated dyes.

Even after conjugation to antibodies, the pHAb dyes retain their fluorescence responsiveness to changes in pH

Although antibody labeling is their primary application, pHAb dyes can conjugate to any protein containing primary amines on lysine residues or thiol groups on cysteine residues.

Here are some recommended instrument parameters: 
• Microplate Reader: Tecan Infinite® M1000 Pro (Ex: 532 nm / Em: 560 nm) 
• Flow Cytometer: Millipore Guava easyCyte 8HT, using yellow fluorescence parameters (Em filter: 583/26) 
• Fluorescence Microscope: Olympus FV 500; TAMRA setting (Ex: 543 nm / Em: 565 nm) 
• Fluorescence Imaging System: Life Technologies EVOS® FL Cell Imaging System, RFP red cube filter set (Ex: 531 nm / Em: 593 nm) 
• Fluorescence Gel Scanner: GE Typhoon® 9410; TAMRA setting (Ex: 532 nm / Em: 580 nm)