Product manual
Product use, preparation, operating conditions and storage.
BODIPY 530/550Reactive dye
BODIPY530/550 Succinimidyl Ester
Cat. No.QH-40-4005
Label proteins, peptides and amino-modified molecules by coupling the NHS ester to accessible primary amines.
Packs & quoteNHS
Activated esterR–NH₂
Primary amineCO–NH–R
Amide linkageExcitation534 nm
Emission551 nm
TargetProtein N-termini, lysine residues and introduced primary amines
Preparation and storage
- Storage
- Store at -20°C, protect from light and moisture
Primary-amine coupling
The NHS ester forms an amide bond with a primary amine; hydrolysis competes with coupling. Proteins usually contain multiple accessible sites, and the input ratio is not the final degree of labeling.
Protein and antibody labeling
For samples with accessible primary amines. Check antigen binding, enzyme activity or the required function after purification.
Amino-modified nucleic acids and peptides
Confirm the number and protection state of amino handles, then choose purification and analysis for the intended product.
Filters and excitation leakage
Check filter bandwidths, dichroics and blocking against the actual excitation and emission values. Some BDP products have closely spaced peaks; peak positions alone do not establish excitation leakage or background. Check with unstained and free-dye controls.
Specifications
Product information
- Product name
- BODIPY530/550 Succinimidyl Ester
- Cat. No.
- QH-40-4005
- Product form
- NHS Ester
- Target
- Protein N-termini, lysine residues and introduced primary amines
- Pack sizes
- 1 mg / 5 mg / 10 mg / 25 mg / 100 mg
Preparation and properties
- Excitation wavelength
- 534 nm
- Emission wavelength
- 551 nm
- CAS
- 216961-93-2
- Molar extinction coefficient
- 77,000 M⁻¹ cm⁻¹
- Storage
- Store at -20°C, protect from light and moisture
Spectral information and detection
Product parameters
Use parameters for the exact catalog number when preparing reagents, setting reagent input or calculating results. Request a lot-specific CoA for test results.
Protocol
Buffer
Avoid competing primary amines such as Tris, glycine and free amino acids. Remove carrier proteins such as BSA when they are not intended for labeling. Use a primary-amine-free buffer compatible with the sample.
Dye and input calculation
Prepare the activated ester using a solvent supported for this product. Convert sample and dye amounts to moles using their molecular weights and record the final organic-solvent fraction.
Aqueous dilution and vessel adsorption
Confirm the supported solvent and stock concentration for the catalog number before diluting into a small sample. Check precipitation, vessel staining and transfer loss, matching final solvent fraction and vessel material when comparing conditions.
Exchange the sample buffer
Exchange into a suitable buffer and check concentration, purity, pH and visible aggregation.
Set up a reaction series
Use a small sample to compare dye input ratios. Mix gently, protect from light and record pH, temperature and time. Select conditions from this product’s instructions and sample tolerance.
Purification and analysis
Select desalting, size-exclusion separation, dialysis or chromatography for the sample size. Remove free reagent before measuring concentration, labeling density and function. Residual free dye raises background and biases labeling-density calculations.
Assess the conjugate
Calculate labeling density with appropriate extinction coefficients and absorbance corrections, then select conditions based on background, recovery and function.
Request product documentation
Select a document and enter your phone and email. Include the lot number for a CoA.
BODIPY530/550 Succinimidyl EsterQH-40-4005
Spectra and measurement conditions
Spectra, measurement conditions and detection settings for the exact catalog number.
Certificate of analysis (CoA)
Lot-specific test results. Include the lot number from the vial.
Safety data sheet (SDS)
Handling, storage and emergency response information.
Quality and lot information
Unlabeled sample
Use an unlabeled aliquot from the same preparation to assess changes in function, aggregation and recovery.
Free dye and negative sample
Include free-dye or no-target-handle controls to distinguish conjugation from adsorption. Match exposure, gain or voltage during detection.
Input titration
Compare a molar input series and assess signal, negative background, product recovery and function together.
Composition after purification
Check conjugate and free-dye fractions together. A clear or strongly fluorescent solution still needs suitable chromatography, electrophoresis or mass spectrometry to establish coupling; assess binding or enzyme activity separately for protein conjugates.
Technical articles
Buffer checks before NHS labeling
Tris and glycine consume NHS dye. BSA and gelatin require separation from the target protein; routine buffer exchange is insufficient.
For protein preparation before NHS-ester labeling; check competing amines and carrier proteins.
Antibody DOL calculation and quality assessment
Calculate DOL with absorbance correction, distinguish input ratio from attached dye, and assess recovery and binding.
For purified protein conjugates; use this dye’s absorbance parameters and distinguish input ratio from DOL.
Application support
Include your sample, formulation and detection method when requesting application guidance.
References
BODIPY dyes and their derivatives: syntheses and spectroscopic properties
Loudet A, Burgess K. · Chemical Reviews · 107(11):4891–4932 · 2007
Reviews BODIPY synthesis, derivatization and spectroscopic properties as background for distinguishing members of the dye family.
DOI: 10.1021/cr078381n
Labeling a protein with fluorophores using NHS ester derivitization
Nanda JS, Lorsch JR. · Methods in Enzymology · 536:87–94 · 2014
Uses FAM ester labeling of a protein to explain primary-amine chemistry and labeling-efficiency assessment. It informs NHS workflow design; input and reaction conditions depend on the actual dye and sample.
Fluorescent-labeled antibodies: Balancing functionality and degree of labeling
Vira S, Mekhedov E, Humphrey G, Blank PS. · Analytical Biochemistry · 402(2):146–150 · 2010
Uses FITC-labeled anti-HA antibody to assess changes in labeling density and antigen binding. It informs input-series design and post-purification functional testing; DOL alone is insufficient for selecting a conjugate.
Amine-Reactive BODIPY Dye: Spectral Properties and Application for Protein Labeling
Ksenofontova KV, Kerner AA, Ksenofontov AA, et al. · Molecules · 27(22):7911 · 2022
Studies solvent and protein-conjugation effects on NHS-Ph-BODIPY spectra, informing evaluation of a conjugate in its final sample environment.
Frequently asked questions
Can it directly label unmodified nucleic acids?
Routine NHS coupling requires an accessible primary amine. Confirm a suitable amino modification on the oligonucleotide.
Is a higher degree of labeling better?
Overlabeling can affect binding, solubility or signal. Assess specific signal, background and function after purification.
Can aqueous stock be stored for extended periods?
Activated esters hydrolyze in water. Use only the stock solvent, concentration and storage conditions supported for this product.
Are BDP, X forms and WGA conjugates interchangeable?
Distinguish structure and purpose. X belongs to the specific product’s linker structure; WGA is an already labeled lectin. Use the exact catalog number’s preparation and workflow rather than inferring membrane permeability or equal solubility from the family name.