Most hydrophobic clay minerals, such as clinochlore, are known to cause problems in the recovery of cassiterite. In this study, a new reagent scheme, i.e., sodium oleate (NaOL) as a collector and Al (III) ions as a depressant, for reverse flotation separation of cassiterite and clinochlore was investigated. The flotation performance and interaction mechanism were studied by microflotation tests, adsorption tests, contact angle measurements, and X-ray photoelectron spectroscopy (XPS) analysis. Results of single mineral flotation experiments showed that NaOL had a different flotation performance on cassiterite and clinochlore, and the addition of Al (III) ions could selectively inhibit the floatability of cassiterite. Reverse flotation tests performed on mixed minerals indicated that the separation of cassiterite and clinochlore could be achieved in the presence of NaOL and Al (III) ions. Adsorption experiments demonstrated that Al (III) ions hindered the adsorption of NaOL on cassiterite surfaces but exerted little influence on the adsorption of NaOL on clinochlore surfaces. Results of contact angle measurements indicated that Al (III) ions could impede the hydrophobization process of cassiterite in NaOL solution. XPS results showed that aluminum species were adsorbed onto the cassiterite surfaces through the interaction with O sites.