Rho1D4 tag / 1D4 tag


Rho1D4 Tag Guide

Content:

1. What is the Rho1D4-tag
2. Rho1D4-tag protein purification products
3. Publications using the Rho1D4-tag
4. Rho1D4-tag protein purification in use
5. References
Rho1D4, also called "1D4" is an affinity tag consisting of nine amino acids (T-E-T-S-Q-V-A-P-A). The tag is preferrably added to the C-Terminus of a protein of interest. Agarose Resins and magnetic beads coupled with the matching Rho1D4-antibody are able to purify proteins that are tagged with Rho1D4. Rho1D4 protein purification assays combine the incredible high specificity of other antibody based protein purification with an high protein yield. For more information regarding Rho1D4 we have written this guide to provide you with all necessary information.

Features

Usage Specific binding and purification of Rho1D4-tagged proteins
Specifity Affinity to Rho1D4-tagged proteins
Stability Highly tolerant to detergents
Bead Ligand Rho1D4 antibody
Protein Yield 3-4 mg/ ml resin or MagBeads
Tag Sequence TETSQVAPA
Why Cube Biotech's Rho1D4-tag protein purification products?
There are numerous reasons to choose Cube Biotech. Here are a few.
Rho1D4 Protein Tag Size Size of the tag

902.9 Da
9 amino acids (TETSQVAPA)		 Rho1D4 Protein Yield Protein yield per ml resin

3-4 mg/ml		 Specificity IconSpecificity of interaction

KD: 20 nM (5)		 Rho1D4 Elution condiions Elution conditions

Rho1D4 peptide, low pH, or tag cleavage

Overview of Cube Biotech's Rho1D4 affinity products for protein purification.

Rho1D4 AgaroseRho1D4 CartridgesRho1D4 Magnetic Beads
Rho1D4 agarose resin (40 µm) Rho1D4 agarose (40 µm) Cartridge Rho1D4 MagBeads (30 µm)
Rho1D4 agarose resin (XL sized) Rho1D4 agarose (40 µm) COMPACT Cartridge Rho1D4 MagBeads (XL sized)
UPDATE SEPTEMBER 2020: Rho1D4 starter sets - For all who want to try out the Rho1D4-tag for the first time.
Now on 50% discount!
Starter Set 1 (1 mL agarose + 5 mg peptide) Starter Set 3 (1 x 1 mL Cartridge + 5 mg peptide) Starter Set 2 (1 mL MagBeads + 5 mg peptide)
  Starter Set 4 (1 x 1 mL Compact Cartridge + 5mg peptide)  
  Rho1D4 protein purification support products  
Rho1D4 peptide (for protein elution) Rho1D4-tagged GFP (for positive controls) Rho1D4 antibody [mouse] (for blots)
 
Rho1D4 products by Cube Biotech were used in the following publications:
 

Rho1D4 protein purification in detail

Purifies protein with high specificity and yield
PureCube resins are produced under strict quality guidelines and each batch undergoes quality checks to ensure that the loaded matrix has a high protein capacity. Combined with the specificity of the antibody-epitope interaction, a purification protocol optimized for the target protein can generate elution fractions with exceptionally high yields. Figure 1 shows purified elution fractions of three different membrane proteins, two membrane proteins stabilized in detergents and the third in MSP1D1 nanodisc. The Rho1D4 tagged proteins were expressed in T.ni insect cells or Hek293 cells, respectively and purified on a column containing PureCube Rho1D4 Agarose beads. The Rho1D4 peptide was used as mild eluent. These bands represent membrane proteins corresponding to monomers, tetramers and heterodimers stabilized in nanodisc. Analysis of the oligomeric state as removal of the eluent peptide, can be done with size-exclusion chromatography.

Pure and active membrane proteins
Purification results for membrane proteins purification like this can not be achieved by using more common resins like Ni-NTA. Therefore Rho1D4 is your way of choice here.
Rho1d4 protein purification
Fig. 1: Coomassie blue stains of successful membrane protein purifications after using the Rho1D4-tag.
A: Protein monomer of ~65kDa.
B: Tetrameric protein of ~135 kDA.
C: Heterodimieric protein, both subunits can be seen. The smallest band corresponding to ~ 28 kDa is a MSP1D1 nanodisc protein used for membrane protein stabilization.

References:

  1. Mattle D. et al.(2015). Mammalian Expression, Purification, and Crystallization of Rhodopsin Variants. In: Jastrzebska B. (eds) Rhodopsin. Methods in Molecular Biology, vol 1271. Humana Press, New York, NY
  2. Brosig, A. et al (2019). The Axonal Membrane Protein PRG2 Inhibits PTEN and Directs Growth to Branches. Cell Reports. 29. 2028-2040.e8. 10.1016/j.celrep.2019.10.039.
  3. Ahmed, Chulbul M., et al. "SRD005825 Acts as a Pharmacologic Chaperone of Opsin and Promotes Survival of Photoreceptors in an Animal Model of Autosomal Dominant Retinitis Pigmentosa." Translational Vision Science & Technology 8.6 (2019): 30-30.
  4. Liu, Da-Wei et al. (2018). The Cone Opsin Repertoire of Osteoglossomorph Fishes: Gene Loss in Mormyrid Electric Fish and a Long Wavelength-Sensitive Cone Opsin That Survived 3R. Molecular Biology and Evolution. 10.1093/molbev/msy241.
  5. Kotov, Vadim et al. (2019). High-throughput stability screening for detergent-solubilized membrane proteins. Scientific Reports. 9. 10.1038/s41598-019-46686-8.
  6. Broser, M. et al., (2020) NeoR a near-infrared absorbing rhodopsin. Nat Commun 11, 5682. https://doi.org/10.1038/s41467-020-19375-8
  7. Swonger Koretz. et al., (2021) Characterization of binding kinetics of A2AR to Gαs protein by surface plasmon resonance. Biophysical Journal,. https://doi.org/10.1016/j.bpj.2021.02.032.
 
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Rho Starter Set 1: PureCube Rho1D4 Agarose (1 ml) + Rho1D4 peptide (5 mg) Rho Starter Set 1: PureCube Rho1D4 Agarose (1...
Cube Biotech offers Rho1D4-tag affinity agarose & the Rho1D4 protein elution peptide together as an affordable set.
Article number: 33199
Sales price: €175.00 * €350.00 *
PureCube Rho1D4 Agarose PureCube Rho1D4 Agarose
Our PureCube Rho1D4 agarose for protein purification. Rho1D4antibody agarose beads for maximum purity and specificity.
Article number: 33101
Sales price: From €239.00 *
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Rho1D4 antibody (0.2 mg) Rho1D4 antibody (0.2 mg)
For the detection of Rho1D4 tagged proteins on westernblots. Our Rho1D4 affinity antibody, from mouse!
Article number: 40020
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Rho1D4 peptide Rho1D4 peptide
Ultra-Pure Rho1D4 peptide. | For Rho1D4-tag protein elution from Rho1D4 agarose or MagBeads | Ready for your research.
Article number: 16201
Sales price: From €111.00 *
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Rho Starter Set 4: PureCube Compact Cartridge Rho1D4 (1x1 ml) + Rho1D4 peptide (5 mg) Rho Starter Set 4: PureCube Compact Cartridge...
Cube Biotech offers Rho1D4-tag affinity COMPACT columns & the Rho1D4 protein elution peptide together as an affordable set.
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PureCube Rho1D4 Agarose XL PureCube Rho1D4 Agarose XL
Our PureCube Rho1D4 XL agarose for protein purification. Rho1D4 antibody agarose beads for maximum purity and specificity.
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PureCube Rho1D4 MagBeads PureCube Rho1D4 MagBeads
PureCube magnetic beads with Rho1D4 / 1D4-tag affinity. For highly specific and ~100% pure membrane protein purifications.
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Rho Starter Set 2: PureCube Rho1D4 MagBeads (1 ml) + Rho1D4 peptide (5 mg) Rho Starter Set 2: PureCube Rho1D4 MagBeads (1...
Cube Biotech offers Rho1D4-tag affinity MagBeads & the Rho1D4 protein elution peptide together as an affordable set.
Article number: 33299
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PureCube Rho1D4 Cartridge PureCube Rho1D4 Cartridge
Cube Biotech offers Cartridges / Columns for FPLC based Rho1D4-tag Protein Purification. Pre-Filled with Rho1D4 agarose.
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PureCube Compact Cartridge Rho1D4 PureCube Compact Cartridge Rho1D4
Our PureCube Compact Cartridge for low pressure chromatography (FPLC), packed with 1 ml or 5 ml Rho1D4 affinity agarose resin
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Rho Starter Set 3: PureCube Rho1D4 Cartridge (1x1 ml) + Rho1D4 peptide (5 mg) Rho Starter Set 3: PureCube Rho1D4 Cartridge...
Cube Biotech offers Rho1D4 tag affinity columns & the Rho1D4 protein elution peptide together as an affordable set.
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Sales price: €205.00 * €410.00 *
PureCube Rho1D4 MagBeads XL PureCube Rho1D4 MagBeads XL
PureCube Rho1D4 affinity Magnetic beads XL for highly pure membrane protein purification. | 5% suspension | ~100% purity
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