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ReadiUse™ Preactivated APC-iFluor® 800 Tandem

Our preactivated APC was premodified with our Buccutite™ FOL (provided). Your antibody (or other proteins) is modified with our Buccutite™ MTA (provided as free sample) to give MTA-modified protein (such as antibody). The MTA-modified protein readily reacts with FOL-modified APC (provided) to give the desired APC-antibody conjugate in much higher yield than the SMCC chemistry. In addition our preactivated APC reacts with MTA-modified biopolymers at much lower concentrations than the SMCC chemistry.
Our preactivated APC was premodified with our Buccutite™ FOL (provided). Your antibody (or other proteins) is modified with our Buccutite™ MTA (provided as free sample) to give MTA-modified protein (such as antibody). The MTA-modified protein readily reacts with FOL-modified APC (provided) to give the desired APC-antibody conjugate in much higher yield than the SMCC chemistry. In addition our preactivated APC reacts with MTA-modified biopolymers at much lower concentrations than the SMCC chemistry.
Ordering information
Price ()
Catalog Number2572
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Spectral properties
Absorbance (nm)804
Extinction coefficient (cm -1 M -1)700000
Excitation (nm)546
Emission (nm)806
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501
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OverviewpdfSDSpdfProtocol


Absorbance (nm)
804
Extinction coefficient (cm -1 M -1)
700000
Excitation (nm)
546
Emission (nm)
806
Allophycocyanin (APC) is a phycobiliprotein isolated from Spirulina sp., a blue-green alga. Like other phycobiliproteins, APC is fluorescent, with an extremely high absorptivity and a high quantum efficiency. Our APC-iFluor® 800 Tandem is a unique fluorophore with an IR emission developed for spectral flow cytometry. Its emission spectrum does not overlap with any of the common fluorophores, making a unique color for multicolor applications in flow cytometry. ReadiUse™ Preactivated APC-iFluor® 800 Tandem is an activated APC protein, and can be easily conjugated to antibodies with much higher conjugation yield than the conventional APC tandems.

Components


A: ReadiUse™ Preactivated APC-iFluor® 800 Tandem1 vial (1 mg)
B: Buccutite™ MTA1 vial (100 µg)
C: Spin Desalting ColumnNot Included

Example protocol


SAMPLE EXPERIMENTAL PROTOCOL

Preparation of pre-activated antibody with Buccutite™ MTA
  1. Reconstitute Buccutite™ MTA in DMSO at ~10 mg/mL.
    Note     Please store unused Buccutite™ MTA at -20 °C and could be used up to two freeze and thaw cycles.
  2. Prepare target antibody (Ab) in pH = 8.5 - 9.0 buffer at concentration above 1 mg/mL.
  3. Add Buccutite™ MTA to Ab solution at the ratio of 8 - 10 µg Buccutite™ MTA/100 µg Ab.
  4. Mix well and react at room temperature for 60 minutes, rotating during the reaction.
  5. Purify the reaction mixture with desalting column to remove unreacted Buccutite™ MTA and exchange buffer to PBS or buffer of your choice.
  6. Collect the Buccutite™ MTA-activated Ab, and estimate the concentration by 70% yield of the original starting amount. 

Conjugation with pre-activated APC-iFluor™ 800 Tandem
  1. Reconstitute pre-activated APC-iFluor™ 800 Tandem in 100 µL ddH2O to 10 mg/mL.
    Note     Reconstituted pre-activated APC-iFluor™ 800 Tandem could be stored at 4 °C for one month, kept from light.
  2. Add APC-iFluor™ 800 Tandem directly to MTA-activated target Ab solution at the ratio of 130 µg APC-iFluor™ 800 Tandem/100 µg MTA-activated Ab.
  3. Rotate the mixture for 60 minutes at room temperature.
  4. The Ab/APC-iFluor™ 800 Tandem conjugates are now ready to use.
    Note      The antibody conjugate should be stored at > 0.5 mg/mL in the presence of a carrier protein (e.g., 0.1% bovine serum albumin) and 0.02% - 0.05% sodium azide. The Ab/APC-iFluor™ 800 Tandem conjugates solution could be stored at 4 °C for up to two months, and kept from light.
  5. (Optional) Ab/APC-iFluor™ 800 Tandem conjugates could be further purified through size exclusion chromatography to get best performance. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Absorbance (nm)804
Extinction coefficient (cm -1 M -1)700000
Excitation (nm)546
Emission (nm)806

Product family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)
ReadiUse™ Preactivated APC-iFluor® 700 Tandem685710700000
ReadiUse™ Preactivated APC-iFluor® 750 Tandem754793700000

References


View all 2 references: Citation Explorer
Flow cytometry APC-tandem dyes are degraded through a cell-dependent mechanism.
Authors: Le Roy, Christine and Varin-Blank, Nadine and Ajchenbaum-Cymbalista, Florence and Letestu, Rémi
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2009): 882-90
Cy7PE and Cy7APC: bright new probes for immunofluorescence.
Authors: Roederer, M and Kantor, A B and Parks, D R and Herzenberg, L A
Journal: Cytometry (1996): 191-7