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iFluor® 770 maleimide

Ordering information
Price ()
Catalog Number1370
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
Physical properties
SolventDMSO
Spectral properties
Absorbance (nm)772
Correction Factor (260 nm)0.09
Correction Factor (280 nm)0.08
Extinction coefficient (cm -1 M -1)2500001
Excitation (nm)777
Emission (nm)797
Quantum yield0.16
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501
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iFluor® 405 goat anti-mouse IgG (H+L)
iFluor® 488 goat anti-mouse IgG (H+L)
iFluor® 514 goat anti-mouse IgG (H+L)
iFluor® 532 goat anti-mouse IgG (H+L)
iFluor® 555 goat anti-mouse IgG (H+L)
iFluor® 594 goat anti-mouse IgG (H+L)
iFluor® 633 goat anti-mouse IgG (H+L)
iFluor® 647 goat anti-mouse IgG (H+L)
iFluor® 680 goat anti-mouse IgG (H+L)
iFluor® 700 goat anti-mouse IgG (H+L)
iFluor® 750 goat anti-mouse IgG (H+L)
iFluor® 790 goat anti-mouse IgG (H+L)
iFluor® 350 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 405 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 488 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 514 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 532 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 555 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 594 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 633 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 647 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 680 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 700 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 750 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 790 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 350 goat anti-rabbit IgG (H+L)
iFluor® 405 goat anti-rabbit IgG (H+L)
iFluor® 488 goat anti-rabbit IgG (H+L)
iFluor® 514 goat anti-rabbit IgG (H+L)
iFluor® 532 goat anti-rabbit IgG (H+L)
iFluor® 555 goat anti-rabbit IgG (H+L)
iFluor® 594 goat anti-rabbit IgG (H+L)
iFluor® 633 goat anti-rabbit IgG (H+L)
iFluor® 647 goat anti-rabbit IgG (H+L)
iFluor® 680 goat anti-rabbit IgG (H+L)
iFluor® 700 goat anti-rabbit IgG (H+L)
iFluor® 750 goat anti-rabbit IgG (H+L)
iFluor® 790 goat anti-rabbit IgG (H+L)
iFluor® 350 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 405 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 488 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 514 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 532 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 555 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 594 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 633 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 647 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 680 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 700 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 750 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 790 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 350-streptavidin conjugate
iFluor® 405-streptavidin conjugate
iFluor® 488-streptavidin conjugate
iFluor® 514-streptavidin conjugate
iFluor® 532-streptavidin conjugate
iFluor® 555-streptavidin conjugate
iFluor® 594-streptavidin conjugate
iFluor® 633-streptavidin conjugate
iFluor® 647-streptavidin conjugate
iFluor® 680-streptavidin conjugate
iFluor® 700-streptavidin conjugate
iFluor® 750-streptavidin conjugate
iFluor 750™ RGD Conjugate
iFluor® 488 azide
iFluor® 405 hydrazide
iFluor® 800 acid
iFluor® 810 acid
iFluor® 820 acid
iFluor® 860 acid
iFluor® 546 goat anti-mouse IgG (H+L)
iFluor® 546 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 568 goat anti-mouse IgG (H+L)
iFluor® 568 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 546 goat anti-rabbit IgG (H+L)
iFluor® 546 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 568 goat anti-rabbit IgG (H+L)
iFluor® 568 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 555 alkyne
iFluor® 555 azide
iFluor® 546-streptavidin conjugate
iFluor® 568-streptavidin conjugate
iFluor® 488 alkyne
iFluor® 350 Styramide *Superior Replacement for Alexa Fluor 350 tyramide*
iFluor® 488 Styramide *Superior Replacement for Alexa Fluor 488 tyramide and Opal 520*
iFluor® 546 Styramide *Superior Replacement for Alexa Fluor 546 tyramide*
iFluor® 555 Styramide *Superior Replacement for Alexa Fluor 555 tyramide and Opal 570*
iFluor® 568 Styramide *Superior Replacement for Alexa Fluor 568 tyramide*
iFluor® 594 Styramide *Superior Replacement for Alexa Fluor 594 tyramide*
iFluor® 647 Styramide *Superior Replacement for Alexa Fluor 647 tyramide*
iFluor® 680 Styramide *Superior Replacement for Alexa Fluor 680 tyramide and Opal 690*
iFluor® 700 Styramide *Superior Replacement for Alexa Fluor 700 tyramide*
iFluor® 750 Styramide *Superior Replacement for Alexa Fluor 750 tyramide*
iFluor® 790 Styramide *Superior Replacement for Alexa Fluor 790 tyramide*
iFluor® 555 Tyramide
iFluor® 647 Tyramide
iFluor® 350 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor® 488 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor® 555 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor® 594 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor® 647 PSA™ Imaging Kit with Goat Anti-Rabbit IgG
iFluor 350™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor 488™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor 555™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor 594™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor 647™ PSA™ Imaging Kit with Goat Anti-Mouse IgG
iFluor® 840 acid
iFluor® 790-streptavidin conjugate
iFluor® 800-streptavidin conjugate
iFluor® 820-streptavidin conjugate
iFluor® 840-streptavidin conjugate
iFluor® 860-streptavidin conjugate
iFluor® 488-Wheat Germ Agglutinin (WGA) Conjugate
iFluor® 555-Wheat Germ Agglutinin (WGA) Conjugate
iFluor® 594-Wheat Germ Agglutinin (WGA) Conjugate
iFluor® 647-Wheat Germ Agglutinin (WGA) Conjugate
iFluor® 488-Concanavalin A Conjugate
iFluor® 555-Concanavalin A Conjugate
iFluor® 594-Concanavalin A Conjugate
iFluor® 647-Concanavalin A Conjugate
iFluor® 350 Tyramide
iFluor® 546 Tyramide
iFluor® 568 Tyramide
iFluor® 594 Tyramide
iFluor® 488 TCO
iFluor® 555 TCO
iFluor® 594 TCO
iFluor® 647 TCO
iFluor® 488 Tetrazine
iFluor® 555 Tetrazine
iFluor® 594 Tetrazine
iFluor® 647 Tetrazine
iFluor® 440-dUTP *1 mM in Tris Buffer (pH 7.5)*
iFluor®488-dUTP *1 mM in Tris Buffer (pH 7.5)*
iFluor® 450 Styramide *Superior Replacement for Opal Polaris 480*
iFluor® 633 tyramide
iFluor® 514 Styramide *Superior Replacement for Opal 540*
iFluor® 532 Styramide
iFluor® 633 Styramide *Superior Replacement for Opal 650*
iFluor® 440 Styramide
iFluor® 460 Styramide
iFluor® 610 Styramide
iFluor® 660 Styramide
iFluor® 405 Styramide
iFluor®488-PEG12-dUTP *1 mM in Tris Buffer (pH 7.5)*
iFluor®555-PEG12-dUTP *1 mM in Tris Buffer (pH 7.5)*
iFluor®647-PEG12-dUTP *1 mM in Tris Buffer (pH 7.5)*
iFluor® 800 goat anti-mouse IgG (H+L)
iFluor® 800 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 810 goat anti-mouse IgG (H+L)
iFluor® 810 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 820 goat anti-mouse IgG (H+L)
iFluor® 820 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 840 goat anti-mouse IgG (H+L)
iFluor® 840 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 860 goat anti-mouse IgG (H+L)
iFluor® 860 goat anti-mouse IgG (H+L) *Cross Adsorbed*
iFluor® 800 goat anti-rabbit IgG (H+L)
iFluor® 800 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 810 goat anti-rabbit IgG (H+L)
iFluor® 810 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 820 goat anti-rabbit IgG (H+L)
iFluor® 820 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 840 goat anti-rabbit IgG (H+L)
iFluor® 840 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 860 goat anti-rabbit IgG (H+L)
iFluor® 860 goat anti-rabbit IgG (H+L) *Cross Adsorbed*
iFluor® 430 Tyramide *Superior Replacement for Opal 480*
iFluor® 450 Tyramide *Superior Replacement for Opal 480*
iFluor® 350 succinimidyl ester
iFluor® 405 succinimidyl ester
iFluor® 488 succinimidyl ester
iFluor® 514 succinimidyl ester
iFluor® 532 succinimidyl ester
iFluor® 555 succinimidyl ester
iFluor® 594 succinimidyl ester
iFluor® 633 succinimidyl ester
iFluor® 647 succinimidyl ester
iFluor® 660 succinimidyl ester
iFluor® 680 succinimidyl ester
iFluor® 700 succinimidyl ester
iFluor® 750 succinimidyl ester
iFluor® 610 succinimidyl ester
iFluor® 710 succinimidyl ester
iFluor® 790 succinimidyl ester
iFluor® 800 succinimidyl ester
iFluor® 810 succinimidyl ester
iFluor® 820 succinimidyl ester
iFluor® 860 succinimidyl ester
iFluor® 546 succinimidyl ester
iFluor® 568 succinimidyl ester
iFluor® 430 succinimidyl ester
iFluor® 450 succinimidyl ester
iFluor® 840 succinimidyl ester
iFluor® 560 succinimidyl ester
iFluor® 670 succinimidyl ester
iFluor® 460 succinimidyl ester
iFluor® 440 succinimidyl ester
iFluor® 665 succinimidyl ester
iFluor® 690 succinimidyl ester
iFluor® Ultra 594 succinimidyl ester
iFluor® Ultra 647 succinimidyl ester
iFluor® Ultra 750 succinimidyl ester
iFluor® 720 succinimidyl ester
iFluor® 740 succinimidyl ester
iFluor® 597 succinimidyl ester
iFluor® 780 succinimidyl ester
iFluor® 570 succinimidyl ester
iFluor® 830 acid
iFluor® 830 succinimidyl ester
iFluor™ 405 azide
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OverviewpdfSDSpdfProtocol


Absorbance (nm)
772
Correction Factor (260 nm)
0.09
Correction Factor (280 nm)
0.08
Extinction coefficient (cm -1 M -1)
2500001
Excitation (nm)
777
Emission (nm)
797
Quantum yield
0.16
AAT Bioquest's iFluor® dyes are optimized for labeling proteins, particularly antibodies. These dyes are bright, photostable, and have minimal quenching on proteins. They can be well excited by the major laser lines of fluorescence instruments (e.g., 350, 355, 405, 488, 555, 633, 638, 647, 660, and 802 nm). iFluor® 770 dyes have fluorescence excitation and emission maxima of ~777 nm and ~797 nm respectively. These spectral characteristics make them a unique color for fluorescence imaging and flow cytometry applications. iFluor® 770 is an excellent acceptor dye for preparing tandem colors with APC and PE. These iFluor® 770 tandem colors offer a set of unique color profiles for spectral flow cytometry. iFluor® 770 is one of the brightest NIR dyes, and some of its antibody conjugates are significantly brighter than those prepared with IRDyes of similar wavelengths, such as IRDye 800RS. iFluor® 770 maleimide is a thiol-reactive form used to conjugate with thiol-containing molecules such as reduced antibodies, thiol-modified oligos, and peptides.

Example protocol


PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

1. iFluor™ 770 maleimide stock solution (Solution B)
Add anhydrous DMSO into the vial of iFluor™ 770 maleimide to make a 10 mM stock solution. Mix well by pipetting or vortex. Note: Prepare the dye stock solution (Solution B) before starting the conjugation. Use promptly. Extended storage of the dye stock solution may reduce the dye activity. Solution B can be stored in freezer for upto 4 weeks when kept from light and moisture. Avoid freeze-thaw cycles.

2. Protein stock solution (Solution A)
Mix 100 µL of a reaction buffer (e.g., 100 mM MES buffer with pH ~6.0) with 900 µL of the target protein solution (e.g. antibody, protein concentration >2 mg/mL if possible) to give 1 mL protein labeling stock solution. Note: The pH of the protein solution (Solution A) should be 6.5 ± 0.5. Note: Impure antibodies or antibodies stabilized with bovine serum albumin (BSA) or other proteins will not be labeled well. Note: The conjugation efficiency is significantly reduced if the protein concentration is less than 2 mg/mL. For optimal labeling efficiency the final protein concentration range of 2-10 mg/mL is recommended.
Optional: if your protein does not contain a free cysteine, you must treat your protein with DTT or TCEP to generate a thiol group. DTT or TCEP are used for converting a disulfide bond to two free thiol groups. If DTT is used you must remove free DTT by dialysis or gel filtration before conjugating a dye maleimide to your protein. Following is a sample protocol for generating a free thiol group:
  1. Prepare a fresh solution of 1 M DTT (15.4 mg/100 µL) in distilled water.
  2. Make IgG solution in 20 mM DTT: add 20 µL of DTT stock per ml of IgG solution while mixing. Let stand at room temp for 30 minutes without additional mixing (to minimize reoxidation of cysteines to cystines).
  3. Pass the reduced IgG over a filtration column pre-equilibrated with "Exchange Buffer". Collect 0.25 mL fractions off the column.
  4. Determine the protein concentrations and pool the fractions with the majority of the IgG. This can be done either spectrophotometrically or colorimetrically.
  5. Carry out the conjugation as soon as possible after this step (see Sample Experiment Protocol). Note: IgG solutions should be >4 mg/mL for the best results. The antibody should be concentrated if less than 2 mg/mL. Include an extra 10% for losses on the buffer exchange column. Note: The reduction can be carried out in almost any buffers from pH 7-7.5, e.g., MES, phosphate or TRIS buffers. Note: Steps 3 and 4 can be replaced by dialysis. 

SAMPLE EXPERIMENTAL PROTOCOL

This labeling protocol was developed for the conjugate of Goat anti-mouse IgG with iFluor™ 770 maleimide. You might need further optimization for your particular proteins. Note: Each protein requires distinct dye/protein ratio, which also depends on the properties of dyes. Over labeling of a protein could detrimentally affects its binding affinity while the protein conjugates of low dye/protein ratio gives reduced sensitivity.

Run conjugation reaction
  1. Use 10:1 molar ratio of Solution B (dye)/Solution A (protein) as the starting point:  Add 5 µL of the dye stock solution (Solution B, assuming the dye stock solution is 10 mM) into the vial of the protein solution (95 µL of Solution A) with effective shaking. The concentration of the protein is ~0.05 mM assuming the protein concentration is 10 mg/mL and the molecular weight of the protein is ~200KD. Note: We recommend to use 10:1 molar ratio of Solution B (dye)/Solution A (protein). If it is too less or too high, determine the optimal dye/protein ratio at 5:1, 15:1 and 20:1 respectively.
  2. Continue to rotate or shake the reaction mixture at room temperature for 30-60 minutes. 

Purify the conjugation
The following protocol is an example of dye-protein conjugate purification by using a Sephadex G-25 column.
  1. Prepare Sephadex G-25 column according to the manufacture instruction.
  2. Load the reaction mixture (From "Run conjugation reaction") to the top of the Sephadex G-25 column.
  3. Add PBS (pH 7.2-7.4) as soon as the sample runs just below the top resin surface.
  4. Add more PBS (pH 7.2-7.4) to the desired sample to complete the column purification. Combine the fractions that contain the desired dye-protein conjugate. Note: For immediate use, the dye-protein conjugate need be diluted with staining buffer, and aliquoted for multiple uses. Note: For longer term storage, dye-protein conjugate solution need be concentrated or freeze dried. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Absorbance (nm)772
Correction Factor (260 nm)0.09
Correction Factor (280 nm)0.08
Extinction coefficient (cm -1 M -1)2500001
Excitation (nm)777
Emission (nm)797
Quantum yield0.16

Product family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
iFluor® 350 maleimide3454502000010.9510.830.23
iFluor® 488 maleimide4915167500010.910.210.11
iFluor® 555 maleimide55757010000010.6410.230.14
iFluor® 647 maleimide65667025000010.2510.030.03
iFluor® 680 maleimide68470122000010.2310.0970.094
iFluor® 700 maleimide69071322000010.2310.090.04
iFluor® 750 maleimide75777927500010.1210.0440.039
iFluor® 790 maleimide78781225000010.1310.10.09
iFluor® 800 maleimide80182025000010.1110.030.08
iFluor® 810 maleimide81182225000010.0510.090.15
iFluor® 820 maleimide8228502500001-0.110.16
iFluor® 860 maleimide8538782500001-0.10.14
iFluor® 532 maleimide5375609000010.6810.260.16
iFluor® 594 maleimide58860418000010.5310.050.04
iFluor® 405 maleimide4034273700010.9110.480.77
iFluor® 430 maleimide4334984000010.7810.680.3
iFluor® 568 maleimide56858710000010.5710.340.15
iFluor® 633 maleimide64065425000010.2910.0620.044
iFluor® 450 maleimide4515024000010.8210.450.27
iFluor® 460 maleimide468493800001~0.810.980.46
iFluor® 665 maleimide667692110,00010.2210.120.09
iFluor® 546 maleimide54155710000010.6710.250.15
iFluor® 840 maleimide8368792000001-0.20.09
iFluor® 780 maleimide78480825000010.1610.130.12
iFluor® 830 maleimide830867----
iFluor® 514 maleimide5115277500010.8310.2650.116
iFluor® 660 maleimide66367825000010.2610.070.08
iFluor® 670 maleimide67168220000010.5510.030.033
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References


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Two-photon peak molecular brightness spectra reveal long-wavelength enhancements of multiplexed imaging depth and photostability.
Authors: Lang, Ryan T and Spring, Bryan Q
Journal: Biomedical optics express (2021): 5909-5919
Preliminary study on the application of en bloc resection combined with near-infrared molecular imaging technique in the diagnosis and treatment of bladder cancer.
Authors: Yang, Yongjun and Yang, Xiaofeng and Liu, Chao and Li, Jiawei
Journal: World journal of urology (2020)
Enhanced broadband fluorescence detection of nucleic acids using multipolar gap-plasmons on biomimetic Au metasurfaces.
Authors: Narasimhan, Vinayak and Siddique, Radwanul Hasan and Hoffmann, Magnus and Kumar, Shailabh and Choo, Hyuck
Journal: Nanoscale (2019): 13750-13757
Effect of Liposomes with Different Double Arms Polyethyleneglycol on Hepatic Metastasis Model Mice and Evaluation Using a Fluorescent Imaging Device.
Authors: Sugiyama, Ikumi and Oikawa, Hiroki and Masuda, Tomoyuki and Sadzuka, Yasuyuki
Journal: Current drug delivery (2017): 668-675
Au nanostructures by colloidal lithography: from quenching to extensive fluorescence enhancement.
Authors: Xie, Fang and Centeno, Anthony and Ryan, Mary R and Riley, D Jason and Alford, Neil M
Journal: Journal of materials chemistry. B (2013): 536-543