Anti Streptavidin

Western Blot of Rabbit Anti Streptavidin Antibody. Lane 1: Streptavidin Reduced (p/n S000 01) [0.05ug]. Lane 2: Opal Prestained Molecular Weight Marker (p/n MB 210 0500). Lane 3: Streptavidin Non Reduced (p/n S000 01) [0.05ug]. Primary Antibody: Rabbit Anti Streptavidin at 1:1000 overnight at 2 8C. Secondary Antibody: Goat Anti Rabbit IgG Peroxidase (p/n 611 103 122) at 1:70,000 for 30 mins at RT. Block: Blocking Buffer for Fluorescent Western Blotting (p/n MB 070) for 60 mins at RT. Expect: ~13kDa reduced, ~60kDa non reduced.

Rockland Rabbit anti Streptavidin (200-4195 lot 23495) and Biotin conjugated Rabbit anti-trypsin inhibitor antibody (200-4679 lot 6594) were used to detect target proteins Trypsin Inhibitor (left) and Streptavidin (right) under reducing (R) and non-reducing (NR) conditions. Reduced samples of purified target proteins contained 4% BME and were boiled for 5 minutes. Samples of ~1ug of protein per lane were run by SDS-PAGE. Protein was transferred to nitrocellulose and probed with 1:1000 dilution of primary antibody (ON 4 C). Detection shown was using Dylight 649 conjugated Donkey anti rabbit (611-743-127 lot 20831 1:10K 1.5 hr RT in MB-070) and imaged on the BioRad VersaDoc System.

Dot Blot showing the detection of Streptavidin. A three-fold serial dilution of DyLightTM 488 Conjugated Streptavidin starting at 200ng was spotted onto 0.45 um nitrocellulose and blocked in 1% BSA-TTBS (p/n MB-013, diluted to 1X) 30 min at 20C. Anti-Streptavidin (RABBIT) Antibody (p/n 200-401-095) was incubated in Blocking Buffer for Fluorescent Western Blotting (p/n MB-070) at 1:1,000 for 1 Hour at 20C. An HRP Gt-a-Rb secondary antibody (p/n 611-103-122) was incubated at 1:40,000 for 30 min at 20C and imaged using the Bio-Rad VersaDocR 4000 MP.

Western Blot results of Rabbit Anti-Streptavidin Antibody. Lane 1: Opal Prestained Molecular Weight Ladder (p/n MB-210-0500). Lane 2: Streptavidin (p/n S000-01). Load: 50ng. Primary Antibody: Rabbit Anti-Streptavidin at 1:1000 overnight at 4C. Secondary Antibody: Goat anti-Rabbit HRP (p/n 611-103-122) at 1:70,000 for 30min at RT. Blocking: BlockOut (p/n MB-073) for 30 min at RT.