ATTO 488 streptavidin 

ATTO 488 streptavidin is a fluorescent reagent that specifically targets and binds to biotin (biotin), formed by the conjugation of ATTO 488 with streptavidin (HY-P3152). ATTO 488 streptavidin enables visualization of the activity of streptavidin immobilized on the surface of polymeric nanoparticles, or acts as a fluorescent probe to detect the selective binding and internalization process of anti-HB-EGF/NA with cells expressing HB-EGF (with no such effect on cells that do not express this receptor). ATTO 488 streptavidin effectively verifies the function of streptavidin conjugated to the surface of nanoparticles and is suitable for research related to atherosclerotic cardiovascular diseases^[1][2][3].

Guidelines (The following is our recommended experimental protocol. This protocol serves only as a reference guide, and specific operations should be adjusted according to your actual needs).
ATTO 488 is used as the fluorescent label for the anti-HB-EGF/NA conjugate. Its main applications include confocal microscopy analysis and flow cytometry (FACS) to track the cellular uptake and intracellular localization of the conjugate in Vero cells (including Vero-H cells stably expressing human HB-EGF and non-expressing Vero cells)^[1].
1. Preparation of Working Solution
1.1 Preparation of labeled conjugate:
Immobilize anti-HB-EGF/NA on Protein G Sepharose gel microspheres.
Subsequently, add biotinylated fluorescent dyes (ATTO 488 or ATTO 700) to the microspheres for the labeling reaction.
After unreacted molecules are eluted and removed, the fluorescently labeled anti-HB-EGF/NA conjugate can be eluted and recovered.
2. Cell Staining Protocol
2.1 Cell Incubation
Mix Vero cells and Vero-H cells with the prepared ATTO 488-labeled anti-HB-EGF/NA conjugate, and incubate at 37°C for 1 hour.
After incubation, wash the cells thoroughly with PBS, followed by further incubation at 37°C for 0 hours or 24 hours.
2.2 Pre-imaging Treatment
For confocal microscopy analysis, further processing of the stained cells is required before observation with a confocal laser scanning microscope:
Mix the cells with Alexa 594-labeled anti-mouse IgG and Hoechst 33342, and incubate on ice for 1 hour.

Notes:
1. Adjust the ATTO 488 incubation time according to actual conditions.
2. This product is for R&D use only, and is not intended for drug, household or other uses.
3. For your safety and health, wear a lab coat and disposable gloves during operation.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

1430816-27-5

[ATTO 488 streptavidin]

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Tsuchida S, et al. Anti-HB-EGF Antibody-Mediated Delivery of siRNA to Atherosclerotic Lesions in Mice. Int Heart J. 2018;59(6):1425-1431.

  [2]. Zhao M, et al. Ultralow- and Low-Background Surfaces for Single-Molecule Localization Microscopy of Multistep Biointerfaces for Single-Molecule Sensing. Langmuir. 2018;34(34):10012-10018.

  [3]. Jo A, et al. Design and Fabrication of Streptavidin-Functionalized, Fluorescently Labeled Polymeric Nanocarriers. Langmuir. 2018;34(51):15783-15794.