Recombinant DIG (DIG45) antibody

• Catalog Number: orb692670
• Category: Antibodies
• Description: Recombinant DIG (DIG45) antibody
• Clonality: Recombinant
• Clone Number: DIG45
• Tested applications: ELISA, WB
• Reactivity: Bacteria
• Concentration: 1.0 mg/mL
• Dilution range: ELISA: 1:2000, WB: 1:1000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-DIG (VHH) Antibody is a recombinant antibody. The clone was isolated from a library prepared from a hyper-immunized llama host and purified by affinity chromatography from bacterial culture. Expect reactivity against DIG-labeled probes and proteins.
• Conjugation: Unconjugated
• Source: E. coli
• Biological Origin: E. coli
• Storage: Store at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: anti-Digoxigenin, Digoxigenin (DIG), 1672-46-4, La
• Note: For research use only
• Application notes: Anti-DIG is a his-tagged monoclonal recombinant antibody designed to detect digoxigenin and its conjugates. This antibody has been tested by ELISA, SDS-PAGE, and western blot and is intended for use in immunological assays including ELISA, western blotting, immunofluorescence and fluorescence activated cell sorting (FACS). Fluorochrome conjugated anti-DIG can be used for detection of sensitive non-radioactive in situ hybridization probes. The antibody can be labeled with dyes, enzymes or fluorescence, directly or secondarily, for visualization and detection of DIG-conjugated molecules by immunoblotting or immunofluorescence. Secondary detection can be achieved using conjugated anti-His tag or anti-VHH antibodies. Optimal titers for applications should be confirmed by the end user.
• Expiration Date: 12 months from date of receipt.

ELISA Results of purified recombinant Anti-DIG Single Domain Antibody reactive against BSA-conjugated DIG. The working dilution for clone DIG45 is 1:2000. Each well was coated in duplicate with 1.0 µg of BSA conjugated DIG (Blue) and BSA only (Purple). The starting dilution of antibody was 45 µg/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 orb344539) and TMB substrate (p/n orb348651).

SDS-PAGE Results of Recombinant Anti-DIG Single Domain Antibody. Lane 1: Anti-DIG-VhH (Clone DIG45) Reduced (1 µg). Lane 2: Opal Prestained Molecular Weight Marker. Lane 3: Anti-DIG-VhH (Clone DIG45) Non-Reduced (1 µg). 4-20% Gel Coomassie Blue stained. Expected MW: ~15 kDa. Observed MW: ~16 kDa.

Western Blot of Anti-DIG Single Domain Antibody. Lane 1: Opal Prestained Molecular Weight Marker (5 µL). Lane 2: DIG-BSA conjugated (0.2 µg). Lane 3: BSA only (0.2 µg). Lane 4: HEK293T lysate (20 µg). Primary Antibody: Anti-DIG (VhH) at 1 µg/mL incubated overnight at 4°C. Secondary Antibody: Mouse Anti-6X HIS DyLight™649 at 1:1000 incubated for 1 hr at RT. Blocking buffer: Universal Buffer (p/n orb348637) for 2 hrs at RT. Expected MW: ~80 kDa in DIG+ BSA. Observed MW: ~80 kDa in DIG+ BSA (arrow); secondary background seen in HEK293T Lysate.