Flow cytometric analysis of Biotin expression on human peripheral blood lymphocytes. Human peripheral blood lymphocytes were stained with Pacific Blue Mouse Anti-Human CD3 Antibody (S0B5590) and either Biotin Rabbit IgG Isotype Control (left panel) or SDT Biotin Rabbit CD19 (S0B80433) antibody (right panel) at 5 μL/test, followed by staining with SDT Anti-Biotin Recombinant Mouse mAb (FITC Conjugate) at 5 μL/test. Flow cytometry and data analysis were performed using an Agilent NovoCyte Quanteon and FlowJo™ software.
Product Details
Product Details
Product Specification
| Host | Mouse |
| Antigen | Biotin |
| Clone Number | S-5555 |
| Antibody Type | Mouse mAb |
| Isotype | IgG1 |
| Application | FCM |
| Purification | Protein A |
| Concentration | 0.2 mg/ml |
| Conjugation | FITC |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 1% BSA, 0.09% sodium azide |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8 °C as supplied |
Dilution
| application | dilution | species |
| ICFCM | 5μl per million cells in 100ul volume | Species Independent |
| FCM | 5μl per million cells in 100ul volume | Species Independent |
Background
As a labeling agent, the core value of Biotin lies in exploiting the extraordinarily strong non-covalent binding between biotin and avidin or streptavidin (with dissociation constants on the order of 10⁻¹⁵ M): biotin is covalently conjugated to antibodies, nucleic acid probes, or other target molecules, and detection or capture is then achieved using avidin/streptavidin conjugated to enzymes, fluorophores, or colloidal gold, thereby enabling signal amplification and specific tracing. Because the biotin molecule is very small (approximately 244 Da), labeling usually does not significantly interfere with the biological activity or binding capacity of the target molecule. This system is widely used in enzyme-linked immunosorbent assays, immunoblotting, immunohistochemistry, and lateral flow strip tests: in nucleic acid detection strips, for example, amplification products are labeled with both biotin and fluorescein—biotin immobilizes the product on the streptavidin test line, while fluorescein binds colorimetric particles to generate a visible signal.
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