Anti-tetraspanin antibodies were purchased at Immunostep (Salamanca, Spain): anti-CD9 (clone VJ1/20), anti-CD63 (clone Tea 3/18), and anti-CD147 (clone VJ1/9). Anti-mouse IgG, bovine serum albumin (BSA), 1-ethyl-3-[3-dimethylaminopropyl]-carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS) were provided by Sigma-Aldrich (Madrid, Spain). HEPES was purchased from Fisher Scientific.
Some materials for strips were provided by Millipore (Darmstadt, Germany): glass fibre membrane (GFCP001000) used as sample pad, nitrocellulose membrane (HF07504XSS) and backing cards (HF000MC100). Absorbent pad was purchased from Whatman (Piscataway, NJ, USA). The sample buffer used was 10 mM HEPES pH 7.4 with 0.05% Tween-20 and 1% BSA.
To prepare the detection and control lines, an IsoFlow reagent dispensing system was used. A guillotine Fellowes Gamma (Madrid, Spain) was used to cut the strips. In order to quantify the intensity of the test line using reflectance measurements, a portable strip reader ESE Quant LR3 lateral flow system (Qiagen Inc., Hilden, Germany) was used.
Some materials for strips were provided by Millipore (Darmstadt, Germany): glass fibre membrane (GFCP001000) used as sample pad, nitrocellulose membrane (HF07504XSS) and backing cards (HF000MC100). Absorbent pad was purchased from Whatman (Piscataway, NJ, USA). The sample buffer used was 10 mM HEPES pH 7.4 with 0.05% Tween-20 and 1% BSA.
To prepare the detection and control lines, an IsoFlow reagent dispensing system was used. A guillotine Fellowes Gamma (Madrid, Spain) was used to cut the strips. In order to quantify the intensity of the test line using reflectance measurements, a portable strip reader ESE Quant LR3 lateral flow system (Qiagen Inc., Hilden, Germany) was used.