Cell fluorescent dye labeling detection experiment

Cell fluorescent dye labeling detection experiment

Zenon ⑧ labeling technology provides a fast, universal, and reliable experimental method for preparing fluorescently labeled antibodies. This labeling method can utilize various fluorescent dyes, and the starting material can be a very small amount of unpurified (such as the culture supernatant of hybridoma cells) antibodies. The antibodies labeled with Zenon @ labeling technology are applicable to all application fields where direct labeled antibodies can be used. The advantages of Zenon @ tagging technology include:

● 10 minute posting time

Each marker contains 1-20 micrograms of antibody

● Compatible with BSA and other stable proteins

● No purification steps

● Flexible antibody labeling

Zenon ⑧ - How does tagging technology work

Zenon @ labeling technology provides a multifunctional and easy-to-use method for labeling human and mouse IgG1, lgG2a, and lgG2b antibodies, as well as rabbit IgG antibodies. The specially designed Zenon @ fragment only binds to the Fc fragment of the primary antibody, and this fast, non covalent binding method can quickly label a small amount of primary antibody (Figure 1). Zenon @ labeling technology is simple and efficient; The entire labeling process only takes 10 minutes.

After the formation of Zenon @ antibody conjugates, they can be stored stably for future use; Non specific lgG needs to be added before use to block unbound Zenon ⑨ fragments, without the need for column purification. The Zenon @ complex can therefore be directly applied to the sample.

The antibodies labeled with Zenon @ labeling technology exhibit similar fluorescence intensity or enzyme activity to the directly labeled antibody conjugates observed (Figure 2).

Figure 1. The working principle of Zenon @ tagging technology.Comparison of non covalent labeling of Zenon @ antibody with covalent labeling of micro labeling kit, monoclonal labeling kit, and protein labeling kit.

Figure 2. The height of labeled antibodies produced by Zenon @ labeling technology is equivalent to or higher than that of directly labeled antibodiesUse conjugated labeling with phycocyanin (APC) or non conjugated labeling with 1 μ g Zenon ⑨ APC complex to label various lymphatic marker antibodies. Then, the labeled resistance

The body is used for staining human peripheral blood lymphocyte samples. When preparing Zenon @ staining complexes, increasing the ratio of Zenon @ labeling reagent to the primary antibody used can further enhance the brightness of the complex.

Zenon @ tagging technology: final tagging flexibility

When you need to change the color of a marker or detect whether a specific fluorescence is suitable for your application or antibody, Zenon @ labeling technology can provide * labeling flexibility. Widely selectable markers and universal labeling techniques help you easily obtain multi-color application images composed of different color combinations in flow cytometry (Figure 3) and fluorescence imaging (Figures 4 and 5).

Figure 3. Use of Zenon ⑧ labeling technique in flow cytometry.Human peripheral blood leukocytes were stained with the following three antibodies: anti-CD3 mouse IgG1 antibody (A21330) pre labeled with Zenon ⑧ Alexa Fluor @ 647 mouse IgG1 labeling kit (Z25008), anti-CD4 mouse IgG1 antibody (A21334) pre labeled with Zenon ⑨ R phycoerythrin mouse IgG1 labeling kit (Z25055), and pre labeled with Zenon ⑧ Alexa Fluor@488 Mouse lgG2a labeling kit (Z25102) labeled with anti-CD8 mouse lgG2a antibody (A21338). Figures A and B show that distinguishing between orange fluorescence signal and green fluorescence signal or red fluorescence signal and orange fluorescence signal can distinguish cells, proving that Zenon @ labeled antibodies do not interfere with each other in the same sample. The instrument and settings used for analyzing the sample are: Coulter Elite flow cytometer, targeting phycoerythrin and Alexa Fluor@488 The dye uses 488 nm excitation light, and for AlexaFluor ⑧ 647 dye, 633 nm excitation light is used.

Figure 4. Using Zenon @ labeling technique in fluorescence imaging.

Cut a coronal section of mouse hippocampus with a thickness of 14 μ m Stain with anti-a-microtubule primary antibody (A11126) labeled with Zenon ⑧ Alexa Fluor @ 488 mouse gG1 labeling kit (Z25002). Slices were stained with NeuroTrace ⑨ 530/615 red fluorescent Nissl dye (N21482) to observe neuronal cell bodies, and Hoechst 33258 (H1398, H3569, H21491) to observe cell nuclei.

Figure 5. Using Zenon @ labeling technique in fluorescence imaging.

Microtubulin and mitochondrial probe labeled bovine pulmonary artery endothelial cell microtubule protein using Zenon @ Alexa Fluor@488 Detection of anti-a-microtubule protein mouse IgG2b monoclonal antibody using mouse lgG2b labeling kit (Z25202), and detection of mitochondria using Zenon ⑨ Alexa Fluor @ 555 mouse lgG2b labeling kit (Z25205) labeled antioxidant phosphorylase complex V subunit a mouse lgG2b monoclonal antibody (A21350). Stain nucleic acid using DAPI (D1306, D3571, D21490).

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