Col-F Collagen Binding Reagent
- 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).
- Optional: Incubate samples with blocking buffer for 60 minutes.
- Add Col-F to the media/buffer for each appropriate sample.
- Incubate the samples with the dye solutions for 5-60 minutes at 37°C.
- Wash samples twice with PBS (allow samples to incubate at room temp for 5-10 minutes in PBS during each wash step).
- 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.
Unbehau R, Luthringer-Feyerabend B, Willumeit-Römer R. The impact of brain cell metabolism and extracellular matrix on magnesium degradation. Acta Biomaterialia. 2020 Sep 2;S1742-7061(20)30513-4. doi: 10.1016/j.actbio.2020.08.043. Online ahead of print. Full Text
Zhou Z, Qu J, He L, Zhu Y, Yang S, Zhang F, Guo T, Peng H, Chen P, Zhou Y. Stiff matrix instigates type I collagen biogenesis by mammalian cleavage factor I complex-mediated alternative polyadenylation. JCI Insight. 2020 Feb 13;5(3). pii: 133972. doi: 10.1172/jci.insight.133972. Full Text
Rogozhnikov D, O’Brien PJ, Elahipanah S, Yousaf MN. Scaffold Free Bio-orthogonal Assembly of 3-Dimensional Cardiac Tissue via Cell Surface Engineering. Sci Rep. Full text.
Hoyle NP, Seinkmane E, Putker M, Feeney KA, Krogager TP, Chesham JE, Bray LK, Thomas JM, Dunn K, Blaikley J, O’Neil, JS. Circadian actin dynamics drive rhythmic fibroblast mobilization during wound healing. Sci Transl Med. 2017.Nov 8;9(415). pii: eaal2774. doi: 10.1126/scitranslmed.aal2774. Abstract
Question: 1. Can the stain retain for up to 48 hours without fixing the tissue after washing away excess Col-F? 2. Do you have the same dye for RED color (not green fluorescent)?
Answer: We recommend that this product be used on fresh or frozen tissue samples, but not on formalinfixed paraffin-embedded samples. We have not gotten very good results when working with fixed and embedded tissues. Our product data sheet further describes our recommendations. In terms of Col-F staining for longer periods, such as up to 48 hours, we have not evaluated this experimentally However, we would expect the staining to be retained so long as the tissue is protected from light and tissue itself is remains stable and has not begun to break down. We do not currently have a product available in red fluorescence, though we may look at adding it in the future.