HighSpec Rho1D4 Agarose resin is a suspension of 40 µm agarose beads, coupled to an anti-Rho1D4 antibody. It is meant for protein purification assays of the highest possible purity, exploiting the specificity of an antibody to its antigen.
- Usage: Specific binding and purification of Rho1D4-tagged proteins
- Specificity: Affinity to Rho1D4-tagged (TETSQVAPA) proteins
- Binding capacity: >3 mg/ml
- Bead Ligand: anti-Rho1D4 antibody
- Bead size: 100 μm
- Matrix: 6% cross-linked agarose
- Recommended flow rates: 0.25 to 1 ml/min
- Max. flow rates: 6 ml/min
- Filling quantity: Delivered as a 50% suspension
- Short-term storage: In neutral buffer at 4 °C
- Long-term storage: In neutral buffer with 20% ethanol at 4 °C
- Required equipment:
- Lysis Buffer
- Wash Buffer
- Elution Buffer (incl. Rho1D4 peptide)
- Ice bath
- Refrigerated centrifuge for 50 ml tube (min 10000×g)
- 50 ml centrifuge tube
- Micropipettor and Micropipetting tips
- Disposable gravity flow columns with capped bottom outlet, 2 ml
- pH meter
- End-over-end shaker
- SDS-PAGE buffers, reagents and equipment Optional: Western Blot reagents and equipment
Protein affinity epitope tags find use in numerous applications in life sciences. Ranging from Western Blot detection, protein localization, ELISA assays, and most importantly protein purification. The possible uses for epitope tags are numerous (Molday and Molday 2014). Rho1D4, sometimes also just called '1D4-tag', is an epitope tag that originates from the C-terminal end of bovine rhodopsin. Its nine amino acids long sequence is TETSQVAPA, which gives it a size of 902.9 Dalton. The affinity tag and the specific monoclonal antibodies were discovered in 1984 by Hodges et al. and MacKenzie et al.
