igg conjugated to beta galactosidase  (SouthernBiotech)

Journal: Viruses

Article Title: The Susceptibilities of Respiratory Syncytial Virus to Nucleolin Receptor Blocking and Antibody Neutralization Are Dependent upon the Method of Virus Purification

doi: 10.3390/v9080207

Figure Lengend Snippet: Infectivity and constituents of RSV stocks. ( A ) RSV was titrated on HeLa cells, fixed, and stained for RSV protein 20 h later. RSV infection was detected with a polyclonal anti-RSV antibody followed by a secondary antibody conjugated to β-galactosidase. The insoluble blue color is a product of X-Gal substrate cleavage by β-galactosidase and indicates RSV infection; ( B ) RSV focus-forming units (FFU blue spots) were manually counted and the titre of each virus preparation (outlined in A) in FFU/mL is shown; ( C , D ) Viral RNA was extracted from RSV stock preparations and then measured by qRT-PCR; ( C ) Equivalent amounts of total RNA for each RSV preparation were probed and amplified. The proportion RSV genome detected is represented and compared to sucrose-purified RSV; ( D ) Viral titres for each of the RSV purification preparations were known and represented as FFU per picogram of total RNA; ( E , F ) RSV particles were harvested from HeLa cells in conditioned media and PEG-6000 precipitated for further purification as outlined in A; ( E ) Coomassie blue staining of RSV stock preparations. The relative bovine serum albumin (BSA) band intensity was measured by densitometry of the 65 kDa band; ( F ) RSV proteins and superoxide dismutase 1 (SOD1) were detected by Western blot; ( G ) CD63 was detected by Western blot as an exosomal protein marker. MW: molecular weight.

Article Snippet: Anti-goat IgG H&L conjugated to β-Galactosidase; cat. No. ab136712 (Abcam Inc., Toronto, ON, Canada), Anti-superoxide dismutase 1; cat. No. sc-11407 (Santa Cruz Biotechnology Inc., Mississauga, ON, Canada), Anti-RSV 4 clone blend; cat. No. NB100-65217 and Anti-CD63 (H5C6); cat. No. NBP2-42225 (Novus Biologicals, Oakville, ON, Canada), Anti-RSV polyclonal; cat. No. B65860G (Meridian Life Science, Memphis, TN, USA), 6 nm Colloidal Gold-AffiniPure anti-mouse IgG (H&L); cat. No. 715-195-150, Horseradish peroxidase-conjugated light chain specific antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA), Alexa Fluor-488 and -647 conjugated secondary antibodies (Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Infection, Staining, Quantitative RT-PCR, Amplification, Purification, Western Blot, Marker, Molecular Weight