Col-F 膠原蛋白結合試劑

綠色螢光探針 Col-F 對膠原蛋白和彈性蛋白具有親和力。這種無毒的膠原結合探針為新鮮和冷凍動物組織中復雜的膠原和彈性結構的螢光三維成像提供了一種簡單方便的工具。使用螢光顯微鏡進行分析。

Background

Collagens and elastins are primarily synthesized by fibroblasts. These molecules are principal components of extracellular matrices. Once outside the cell, collagen assembles into fibrils and fibers that provide mechanical strength to tissues. Elastin is secreted by cells, and also forms fibers which crosslink to create a flexible network of fibers and sheets. Col-F is a low molecular weight fluorescent probe that exhibits affinity for collagen and elastin. Col-F easily penetrates between cells and into tissues where it can then bind to collagen and elastin fibers via hydrogen bonds and noncovalent hydrophobic interactions. Although Col-F has shown binding affinity for collagen and elastin, because the labeling is non-specific in nature, other tissue types may also be labeled. Analyze samples using a fluorescence microscope.

Reagent Name

COL-F

Target

Collagen and Elastin

Excitation / Emission

490 nm / 515-520 nm

Method of Analysis

Fluorescence microscope

Sample Type

Cell culture, tissue

Storage

-20°C

Country of Origin

United States

Protocols

1. When ready to use, reconstitute the 0.5 mg Col-F vial by adding 100 µL DMSO (this creates a stock at 6.8 mM).

2. Optional: Incubate samples with blocking buffer for 60 minutes.

3. Add Col-F to the media/buffer for each appropriate sample.

4. Incubate the samples with the dye solutions for 5-60 minutes at 37°C.

5. Wash samples twice with PBS (allow samples to incubate at room temp for 5-10 minutes in PBS during each wash step).

6. Once wash steps are complete, mount a coverslip on each slide. Samples are now ready to visualize with a fluorescence microscope capable of excitation at 490 nm, and emission at 515-520 nm.

Citations (5)

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