Application of Antibody Fragments as Crystallization Enhancers

Generation of Recombinant Antibody Fragments for Membrane Protein Crystallization

“…Subtype specific first-strand synthesis is an alternative if amplification of pseudogenes needs to be suppressed (Hunte & M€ unke, 2004). One to five micrograms of total RNA (alternatively 50-500 ng of mRNA) and 40 pmol oligo-dT primer (H 2 O added to a final volume of 11 μl) are heated for 10 min at 70°C and immediately put on ice.…”

Modulation of the Antigenic Peptide Transporter TAP by Recombinant Antibodies Binding to the Last Five Residues of TAP1

“…26 Total RNA of hybridoma cells expressing the mAb148.3 was extracted with the RNAeasy Mini Kit (Qiagen) followed by poly(A)-mRNA isolation with the Oligotex Direct mRNA Mini Kit (Qiagen). The V L and V H chain genes were PCR-amplified using a set of degenerate primers, 44 cloned into the pDrive vector (Invitrogen) and sequenced. To determine genetic heterogeneity, sequences were compared with antibody genes in the Kabat database.…”