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Western blot analysis of Digoxigenin was performed by loading 2 µg of Digoxigenin conjugated to BSA (lane1) and BSA alone (lane2) using Novex® NuPAGE® 12 % Bis-Tris gel (NP0342BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. Digoxigenin was detected at ~60 kDa using ABfinity™ Digoxigenin Recombinant Rabbit Oligoclonal Antibody (710019) at 0.5-1 µg/ml in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Rabbit IgG-HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005). BSA which was used for conjugation was run on a SDS-PAGE and stained with Simply Blue safe stain (LC6065).
|Tested species reactivity||Chem|
|Host / Isotype||Rabbit / IgG|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Western Blot (WB)||0.5-1µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A.
ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Digoxigenin (DIG), a steroid, is a derivative of the flowers and leaves of plants – Digitalis. Due to the high immunigenicity of this small molecule it is highly preferred in many molecular biology applications. DIG can be covalently added to proteins or nucleic acids which makes it very handy in its diverse applications. The digoxigenin markers are detected with anti-digoxigenin antibodies, which may be labeled with dyes or enzymes for visualization and detection.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.