Rho1D4 tag / 1D4 tag


Purification Resins
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

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 a purification run for chemokine receptor 4 (CXCR4). The tagged protein was expressed in E. coli, solubilized with Fos-Choline®-14 and purified on a column containing PureCube Rho1D4 Agarose beads. Using the rho1D4 peptide as eluent, the 4 elution fractions contained a total recoved protein concentration of 0.8, 1.0, 0.85 and 0.6 mg/mL. A western blot shows CXCR4 at approximately 65 kDa and 35 kDa. These bands represent dimers and monomers of the 39.7 kDa membrane protein. Separation of monomers and dimers, as well as removal of the eluent peptide, can be done with size-exclusion chromatography.

Rho1D4_purification_efficiency
Fig. 2: Purification of chemokine receptor 4 (CXCR4) using PureCube Rho1D4 Agarose. Total E.coli lysate (TL) was solubilized with Fos-Choline-14 and the soluble fraction (SF) was incubated on an immunoaffinity column loaded with rho1D4 antibody. Wash fractions (W1-W3) show no detectable loss of target protein. Concentration of eluted CXCR4 in elution fractions (E1-E4) ranged 0.6-1.0 mg/mL as determined spectrophotometrically.
Pure and active membrane proteins
Purification results for membrane proteins purification like this can not be archived 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 sucessfull membrane protein purifications afther using the Rho1D4 tag. A: Protein monomere of ~65kDa. B: Tetrameric protein of ~135 kDA. C: Heterodimieric protein, both subunity can be seen and a smaller n

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.
Rho1D4 tag / 1D4 tag
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PureCube Rho1D4 Agarose PureCube Rho1D4 Agarose
Our PureCube Rho1D4 agarose for 100% pure protein purification. Antibody based beads for maximum purity and specificity.
Article number: 33101
Sales price: From €232.00 *
Rho Starter Set 1: PureCube Rho1D4 Agarose (1 ml) + Rho1D4 peptide (5 mg) Rho Starter Set 1: PureCube Rho1D4 Agarose (1...
Rho1D4 of 2 ml 50% Rho1D4 Agarose and 5 mg lyophilized Rho1D4 peptide. ► 50% discount compared to the individual products
Article number: 33199
Sales price: €171.00 *
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 €107.00 *
Rho1D4 antibody (0.2 mg) Rho1D4 antibody (0.2 mg)
0.2 mg Rho1D4 antibody in solution  
Article number: 40020
Sales price: €279.00 *
PureCube Rho1D4 MagBeads PureCube Rho1D4 MagBeads
1 ml 5% Rho1D4 Magnetic Beads
Article number: 33201
Sales price: From €246.00 *
Rho Starter Set 2: PureCube Rho1D4 MagBeads (1 ml) + Rho1D4 peptide (5 mg) Rho Starter Set 2: PureCube Rho1D4 MagBeads (1...
Set of 1 ml 5% Rho1D4 MagBeads and 5 mg lyophilized Rho1D4 peptide. Provides a 50% discount compared to purchase of individual items.
Article number: 33299
Sales price: €177.00 *
PureCube Rho1D4 Cartridge PureCube Rho1D4 Cartridge
Cartridge for low pressure chromatography (FPLC), packed with 1 ml Rho1D4 agarose  
Article number: 33301
Sales price: From €291.00 *
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
Article number: 33302
Sales price: From €291.00 *
Rho Starter Set 4: PureCube Compact Cartridge Rho1D4 (1x1 ml) + Rho1D4 peptide (5 mg) Rho Starter Set 4: PureCube Compact Cartridge...
Set of 1x 1ml Rho1D4 prepacked Compact Cartridge and 5 mg lyophilized Rho1D4 peptide
Article number: 33398
Sales price: €199.00 *
Rho Starter Set 3: PureCube Rho1D4 Cartridge (1x1 ml) + Rho1D4 peptide (5 mg) Rho Starter Set 3: PureCube Rho1D4 Cartridge...
Set of 1x 1ml Rho1D4 prepacked Cartridge and 5 mg lyophilized Rho1D4 peptide. Provides a 50% discount compared to purchase of individual items.
Article number: 33399
Sales price: €199.00 *
PureCube Rho1D4 Agarose XL PureCube Rho1D4 Agarose XL
2 ml 50% Rho1D4 Affinity Agarose with medium particle size of 400 micrometer, for special applications
Article number: 57601
Sales price: From €277.00 *
PureCube Rho1D4 MagBeads XL PureCube Rho1D4 MagBeads XL
1 ml 5% Rho1D4 Magnetic Beads. Particle size 70-120 micrometer, for special applications.
Article number: 57701
Sales price: From €303.00 *
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