Anti DIG, recombinant VHH

SDS PAGE Results of Recombinant Anti DIG (VhH) Single Domain Antibody. Lane 1: Anti DIG VhH (Clone DIG44) Reduced (1ug). Lane 2: Opal Prestained Molecular Weight Marker (p/n MB 210 0500). Lane 3: Anti DIG VhH (Clone DIG44) Non Reduced (1ug). 4 20% Gel Coomassie Blue stained. Predicted MW: ~15kDa. Observed MW: ~18kDa Reduced, ~16kDa Non Reduced.

Western Blot of Anti-DIG Single Domain Antibody. Lane 1: Opal Prestained Molecular Weight Marker (p/n MB-210-0500) (5μL). Lane 2: DIG-BSA conjugated (0.2μg). Lane 3: BSA only (0.2μg). Lane 4: HEK293T lysate (p/n W09-001-GX5) (20μg). Left Blot - Primary Antibody: Anti-DIG (VhH) at 1μg/mL incubated overnight at 4°C. Right Blot - No Primary. Secondary Only Control. Secondary Antibody: Mouse Anti-6X HIS DyLight？649 (p/n 200-343-382) at 1:1000 incubated for 1hr at RT. Blocking buffer: Universal Buffer (p/n MB-070) for 2hrs at RT. Expected MW: ~80kDa in DIG+BSA. Observed MW: (arrow) ~80kDa in DIG+BSA; secondary background seen in current tested lot of HEK293T Lysate.

ELISA Results of purified recombinant Anti-DIG Single Domain Antibody reactive against BSA-conjugated DIG. The working dilution for clone DIG44 is 1:1000. Each well was coated in duplicate with 1.0ug BSA-conjugated DIG (Red) and BSA only (Green). The starting dilution of antibody was 45μg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using Sodium Carbonate blocking buffer, Mouse Anti-6X HIS Epitope Tag HRP conjugated Antibody (p/n 200-303-382) and TMB substrate (p/n TMBE-1000).