Rose-β-D-Gal

Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)^[1][2].
1. Prepare 25 mg/mL Rose-β-D-Gal stock solution in Dimethyl Sulfoxide (DMSO). It is vital to protect the stock solution from the light.
2. Prepare rinse buffer for Rose-β-D-Gal staining: 0.1% Sodium Deoxycholate, 0.2% IGEPAL CA-630, 2 mM MgCl₂ in 0.1 M Na Phosphate buffer (pH 7.3).
3. Prepare substrate solution for Rose-β-D-Gal fresh, containing 1 mg/mL Rose-β-D-Gal, 5 mM Potassium Ferricyanide, and 5 mM Potassium Ferrocyanide in the rinse buffer.
4. Rehydrate frozen tissue sections in PBS: quickly wash with PBS twice, wash with rinse buffer for 10 min 3 time.
5. Expose to β-galactosidase substrate Rose-β-D-Gal: stain at 37°C for as long as needed to see stain, up to an overnight time period, keep covered and in the dark during color development. Wash with PBS for 5 min twice.
6. Fix tissue in 4% PFA, unless on intend to continue with in situ hybridizations, and continue to step 4.
Note: Rose-β-D-Gal is not very stable in alcohols or organic solvents, so must use water-based counterstains such as Gill's hematoxylin and cover slipping with aqueous mountant.

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