Product manual
Product use, preparation, operating conditions and storage.
Fluorescent proteinsFluorescent protein
APC
Cat. No.QH-50-5003
A fluorescent-protein reagent for developing protein or antibody conjugates and related detection methods.
Packs & quoteCheck protein condition
Small-scale coupling
Resolve conjugate, free protein and aggregates
Excitation652 nm
Emission662 nm
TargetMolecules coupled through a suitable conjugation route
Preparation and storage
- Preparation
- Aqueous formulation
- Storage
- Store at 2–8°C, protect from light. Do not freeze
Fluorescent-protein detection
This reagent provides a fluorescent-protein signal but is not itself a sample-specific antibody. Antibody or other target labeling requires a suitable activation, coupling and purification route that preserves the protein.
Fluorescent-conjugate development
After preparing the target conjugate, assess binding, free fluorescent-protein carryover and aggregation.
Multicolor detection
Configure the detection channel from this reagent’s optical properties, lasers and filters, then establish single-stain references.
Specifications
Product information
- Product name
- APC
- Cat. No.
- QH-50-5003
- Product form
- Fluorescent proteins
- Target
- Molecules coupled through a suitable conjugation route
- Pack sizes
- 1 mg / 5 mg / 10 mg / 25 mg / 100 mg
Preparation and properties
- Excitation wavelength
- 652 nm
- Emission wavelength
- 662 nm
- Molar extinction coefficient
- 730,000 M⁻¹ cm⁻¹
- Preparation and sample
- Aqueous formulation
- Storage
- Store at 2–8°C, protect from light. Do not freeze
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
Aqueous protein handling
Use an appropriate aqueous buffer for the formulation, avoiding organic solvents, harsh mixing and unsuitable freeze–thaw cycles.
Conjugation route
Check existing sites and any linker handles to be introduced. Do not use a fluorescent-protein reagent as though it were a small-molecule NHS dye.
Confirm activation state
Confirm by catalog number whether an activated handle is supplied and plan matching handles on the fluorescent material and target. Follow the supplied group for preactivated products, avoiding reactivation; remove free activator or reductant from each processed component.
Check protein condition
Check concentration, buffer and optical properties, including visible precipitation and aggregation.
Small-scale coupling
Use a validated activation and coupling procedure, titrating inputs and recording recovery and reaction conditions.
Resolve conjugate, free protein and aggregates
Both the fluorescent material and target antibody are macromolecules. Ordinary desalting or ultrafiltration does not ensure separation of the final conjugate from free fluorescent protein. Validate the resolving power of size-exclusion or another method, then assess recovery, purity and function.
Evaluate detection and storage
Assess positive separation, negative background, function and changes after storage of the purified conjugate.
Request product documentation
Select a document and enter your phone and email. Include the lot number for a CoA.
APCQH-50-5003
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
Unconjugated and negative samples
Compare free protein and the target conjugate to assess nonspecific adsorption.
Single-stain reference
Use the actual conjugate for channels and multicolor spillover correction.
Unlabeled sample
Use an unlabeled aliquot from the same preparation to assess changes in function, aggregation and recovery.
References
Phycobiliprotein reagents, activation and conjugate separation
Thermo Fisher Scientific · Phycobiliproteins · MP 00800
Manufacturer guidance for phycobiliproteins such as R-PE/APC, distinguishing native reagents, derivatives and macromolecular conjugate purification. PerCP is not a phycobiliprotein; confirm its activation, purification and storage separately.
Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)
Cossarizza A, Chang HD, Radbruch A, et al. · European Journal of Immunology · 51(12):2708–3145 · 2021
Provides methodological guidance on detection settings, controls and experimental records for downstream conjugate evaluation.
Frequently asked questions
Can it be used as a specific antibody?
It is a fluorescent-protein or tandem reagent. Specific recognition requires a suitable antibody or targeting molecule.
Can it be prepared in DMSO like a small dye?
Fluorescent proteins require structural preservation. Follow the aqueous formulation rather than small reactive-dye stock methods.
Can fixation alter channel behavior?
Fixation, permeabilization and storage can affect signal. Validate the complete process with the actual conjugate.
Should a preactivated reagent be activated again?
First identify the supplied reactive group, then prepare a matching target handle. Reactivation can change handle density and protein condition; the reagent name alone does not establish activation.