LO1 was purified from hybridoma supernatants by affinity chromatography on protein G sepharose and when needed using a specific anti-idiotype column. An ELISA was used to confirm functional reactivity of LO1 before and after labeling, using MDA-LDL or H3 (anti-LO1 idiotype) on the solid phase, as previously21 (link). Antibodies were labeled with the NIRF fluorochrome VivoTag-S 750-MAL (PerkinElmer, Massachusetts USA) and retention of function confirmed by ELISA. Further details are in the supplementary section.
Vivotag s 750 mal
Manufactured by PerkinElmer
Sourced in United States
VivoTag-S 750-MAL is a fluorescent dye used for in vivo imaging. It is a near-infrared dye with a maximum excitation wavelength of 749 nm and a maximum emission wavelength of 772 nm. This dye can be used to label biomolecules for detection in small animal models.
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Lab products found in correlation
2 protocols using vivotag s 750 mal
1
Antibody Labeling and Functional Validation
2
Fluorescent Antibody Conjugation Protocol
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Conjugation of 78Fc or control human IgG (Genscript) with VivoTag-S 750 and VivoTag-S 750MAL (Perkin Elmer) was performed according to the manufacturer's instructions. Briefly, 1 mL of a 1 mg/mL antibody solution in conjugation buffer (50mM carbonate/bicarbonate buffer, pH8.5) was mixed with 30μL of VivoTag-S 750 at a concentration of 10mg/mL in Dimethylsulfoxide (Sigma) and then incubated at room temperature for 1 h. After dialysis in 1×PBS, the reaction mixture was filtered through a 0.2 μm syringe filter. Aliquots of labeled protein were stored at 4 °C in the dark until ready to analyze.
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