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Biotin labelled bsa

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Biotin-labelled Bovine Serum Albumin (Biotin-labelled BSA) is a protein-based reagent used in various bioanalytical techniques. It consists of the albumin protein from bovine serum that has been covalently modified with biotin molecules. The biotin label allows for the detection and capture of the BSA protein through interactions with streptavidin or avidin-based systems.

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Lab products found in correlation

Cm5 sensor chip, by GE Healthcare (2 mentions) Biacore t100 instrument, by GE Healthcare (2 mentions)

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BSA-biotin (16 citations)

2 protocols using biotin labelled bsa

1

Biotinylated SID Constructs for SPR

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The M. smegmatis SID constructs (wildtype and 3xE mutant) were expressed in the pBXNH3A vector, which allows for N-terminal biotinylation via the Avi-tag49 (link). Expression and purification were performed as described above. Biotinylation was performed simultaneously with 3C protease cleavage overnight. For biotinylation, 5 mM ATP, 10 mM MgOAc, 1.2-fold molar excess of biotin and 16 μg/ml BirA were added. The biotinylated SID constructs were used for SPR measurement using a Biacore T100 instrument (GE Healthcare). Approximately 15’000 response units of the SID constructs were coated onto a Sensor Chip CM5 (GE Healthcare). Afterwards, the coated lanes were saturated by coating with biotin-labelled BSA (Sigma) to avoid unspecific binding of cMBT to the surface of the chip. HPLC purified Fe-cMBT (MW 799 Da) was used as analyte at the concentrations of 0, 6.25, 12.5, 25, 50 and 100 μM. Measurements were conducted as technical duplicates.
Arnold F.M., Weber M.S., Gonda I., Gallenito M.J., Adenau S., Egloff P., Zimmermann I., Hutter C.A., Hürlimann L.M., Peters E., Piel J., Meloni G., Medalia O, & Seeger M.A. (2020). The ABC exporter IrtAB imports and reduces mycobacterial siderophores. Nature, 580(7803), 413-417.
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2

Biotinylated SID Constructs for SPR

Check if the same lab product or an alternative is used in the 5 most similar protocols
The M. smegmatis SID constructs (wildtype and 3xE mutant) were expressed in the pBXNH3A vector, which allows for N-terminal biotinylation via the Avi-tag49 (link). Expression and purification were performed as described above. Biotinylation was performed simultaneously with 3C protease cleavage overnight. For biotinylation, 5 mM ATP, 10 mM MgOAc, 1.2-fold molar excess of biotin and 16 μg/ml BirA were added. The biotinylated SID constructs were used for SPR measurement using a Biacore T100 instrument (GE Healthcare). Approximately 15’000 response units of the SID constructs were coated onto a Sensor Chip CM5 (GE Healthcare). Afterwards, the coated lanes were saturated by coating with biotin-labelled BSA (Sigma) to avoid unspecific binding of cMBT to the surface of the chip. HPLC purified Fe-cMBT (MW 799 Da) was used as analyte at the concentrations of 0, 6.25, 12.5, 25, 50 and 100 μM. Measurements were conducted as technical duplicates.
Arnold F.M., Weber M.S., Gonda I., Gallenito M.J., Adenau S., Egloff P., Zimmermann I., Hutter C.A., Hürlimann L.M., Peters E., Piel J., Meloni G., Medalia O, & Seeger M.A. (2020). The ABC exporter IrtAB imports and reduces mycobacterial siderophores. Nature, 580(7803), 413-417.
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