Goat anti human secondary antibody

Manufactured by Thermo Fisher Scientific

Goat anti-human secondary antibody is a laboratory reagent used to detect and identify human primary antibodies in immunoassays. It binds to the Fc region of human immunoglobulins, allowing for the amplification and visualization of the target antibody signal.

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Lab products found in correlation

Flow cytometry staining buffer solution, by Thermo Fisher Scientific (2 mentions) Lsrfortessa cell analyzer, by BD (2 mentions) Nhs fitc, by Thermo Fisher Scientific (1 mentions)

2 protocols using goat anti human secondary antibody

Cellular Affinity Determination Protocols

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Indirect and direct methods were used to test the cellular affinity according to previous protocols.^[¹⁹^] Briefly, for indirect method, lymphoma cells were incubated with dara (5 × 10⁻⁹m) and then a goat anti‐human secondary antibody (3 µg mL⁻¹, ThermoFisher Scientific) in Flow Cytometry Staining Buffer Solution (eBioscience) at the concentration of 1 × 10⁶ cells mL⁻¹. For direct method,
N‐Hydroxysuccinimide –fluorescein (NHS–FITC, ThermoFisher)‐conjugated antibody was prepared and purified.^[²⁵^] Then, FITC‐conjugated DTPA– or Df–dara were tested and processed using a LSRFortessa cell analyzer (BD Biosciences) and FlowJo software (Tree Star) for the mean fluorescence intensities.

Kang L., Li C., Rosenkrans Z.T., Huo N., Chen Z., Ehlerding E.B., Huo Y., Ferreira C.A., Barnhart T.E., Engle J.W., Wang R., Jiang D., Xu X, & Cai W. (2021). CD38‐Targeted Theranostics of Lymphoma with 89Zr/177Lu‐Labeled Daratumumab. Advanced Science, 8(10), 2001879.

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Daratumumab Binding Assay by Flow Cytometry

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Cells were washed with cold PBS buffer and incubated with Flow Cytometry Staining Buffer Solution (eBioscience, USA) at a concentration of 1 × 10⁶ cells/mL at RT for 1 h. Cells were then incubated with 200 µL of daratumumab (5 or 25 µg/mL in abovementioned buffer) on ice for 60 min and then washed with cold PBS for three times. Cells were incubated with a goat anti-human secondary antibody (3 µg/mL, ThermoFisher Scientific) on ice for 30 min and washed again. The cells were analyzed using a LSRFortessa cell analyzer (BD Biosciences) and mean fluorescence intensities were processed using the FlowJo software (Tree Star, USA).

Kang L., Jiang D., England C.G., Barnhart T.E., Yu B., Rosenkrans Z.T., Wang R., Engle J.W., Xu X., Huang P, & Cai W. (2018). ImmunoPET imaging of CD38 in murine lymphoma models using 89Zr-labeled daratumumab. European journal of nuclear medicine and molecular imaging, 45(8), 1372-1381.

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