Adeno-Associated Virus-Based Vectors - Viral Vector Approaches in Neurobiology and Brain Diseases - page 31

Storm phosphorImager (GE Healthcare, Storm 860) and
ImageQuant densitometry software.
(b) ELISA
AAV2 titration ELISA kit (Progen, cat. no. PRATV).
Incubator at 37 °C.
Microplate absorbance reader (450 nm) (Bio-Rad, iMark, cat.
no. 168-1135)
(c) Infectious center assay
C12 cell line [ 59 ].
Adenovirus serotype 5 (Ad5).
DMEM with glutamax and sodium pyruvate supplemented with
10 % HI-FBS, 1× penicillin-streptomycin. 1× trypsin.
1× PBS.
Denaturing solution: 1.5 M NaCl, 0.5 M NaOH.
Neutralizing solution: 0.5 M NaCl, 0.5 M Tris-HCl, pH 7.5.
2× SSC: 0.3 M NaCl, 30 mM sodium citrate, pH 7.0.
DNA nylon membrane filter, 25 mm (Millipore, cat. no.
ANWP02500)
3MM chr blotting paper (Whatman).
96-well plate for tissue culture (Becton Dickinson, cat. no.
734-0019).
Kodak BioMax MR film (Carestream Health, cat. no. 873 6936).
1225 Sampling manifold (Millipore, cat. no. XX2702550).
X-ray film developer.
(d) Transduction assay
HeLa cells (ATCC, cat. no. CCL-2).
DMEM with glutamax and sodium pyruvate supplemented with
10 % HI-FBS, 1× penicillin-streptomycin. 1× trypsin.
24-well plate for tissue culture (Becton Dickinson, cat. no.
734-0020).
1× PBS.
FACS tube (Becton Dickinson, cat. no. 352008).
FACScalibur and CellQuest software (Becton-Dickinson)

(a) Assay for protein purity
6× Laemmli buffer.
10 % SDS-polyacrylamide gel.
1× Tris-Glycine, 0.1 % SDS.
Dual color protein ladder (Bio-Rad, cat. no. 161-0374).
Krypton™ Infrared Protein Stain (Pierce, cat. no. 53071).

2.5 Vector Quality
Control