Talon affinity chromatography

Manufactured by Takara Bio

Sourced in Canada, United States, United Kingdom

TALON affinity chromatography is a specialized lab equipment used for protein purification. It utilizes a cobalt-based resin to selectively bind and capture histidine-tagged proteins. The core function of this equipment is to enable efficient and reliable purification of target proteins from complex mixtures.

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Lab products found in correlation

Protease inhibitor cocktail, by Roche (1 mentions)

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TALON metal affinity chromatography (12 citations) TALON chromatography (2 citations) TALON affinity chromatography resin (2 citations) Talon Immobilized Metal Affinity Chromatography (IMAC) (2 citations)

4 protocols using talon affinity chromatography

Recombinant P. falciparum IspC Purification

Cited 3 times

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The cloning, expression, and purification of P. falciparum IspC were performed, as described previously.^{10 (link),12 (link),17 (link)} Briefly, the P.
falciparum IspC gene was cloned into a pET100/D-TOPO
vector to facilitate the expression of an N-terminal His₆-tagged protein. The recombinant protein was expressed in Escherichia coli Rosetta2(DE3) cells obtained from
Novagen (San Diego, CA). E. coli was
cultured at 37 °C in Luria-Bertani media supplemented with 100
μg/mL ampicillin and 34 μg/mL chloramphenicol with constant
shaking at 250 rpm. Agar (1.5% w/v) was added to prepare solid media.
Protein was isolated and purified from the cells via chemical lysis
and TALON affinity chromatography (Clontech Laboratories, Mountain
View, CA).

Girma M.B., Ball H.S., Wang X., Brothers R.C., Jackson E.R., Meyers M.J., Dowd C.S, & Couch R.D. (2021). Mechanism of Action of N-Acyl and N-Alkoxy Fosmidomycin Analogs: Mono- and Bisubstrate Inhibition of IspC from Plasmodium falciparum, a Causative Agent of Malaria. ACS Omega, 6(42), 27630-27639.

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Purification of His-tagged TSR1 Domain

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Bacterial codon optimized gene encoding His-tagged TSR1 (Alanine₄₈-Leucine₁₂₆) domain was synthesized (Genscript) and sub-cloned into pET 100/D-TOPO vector. The construct was freshly transformed into competent Origami 2 E.coli cells (Novagen). Cells were grown to an OD600 of 0.6, chilled on ice and 0.2 mM IPTG added. The cells were cultured overnight in a shaker incubator (220 rpm) at 18 °C. Cells were harvested (6500×g at 4 °C). The resulting frozen pellet was lysed in chilled 50 mM Tris-HCl pH 8, 0.5 M NaCl, 10% glycerol, 0.1% Triton X-100, protease inhibitor cocktail (Roche) containing 100 μg/ml lysozyme for 30 min on ice. Resuspended cells were sonicated 6 × 20 s bursts at 12% setting with cooling in between and then clarified by centrifugation at 10,000×g for 20 min at 4 °C. Clarified lysate was then purified by TALON affinity chromatography (Takara, ClonTech) using an imidazole wash step (10 mM) and then eluted in PBS buffer containing 300 mM imidazole. Protein was further purified on Superdex 75 column run in PBS buffer.

Fresquet M., Rhoden S.J., Jowitt T.A., McKenzie E.A., Roberts I., Lennon R, & Brenchley P.E. (2020). Autoantigens PLA2R and THSD7A in membranous nephropathy share a common epitope motif in the N-terminal domain. Journal of Autoimmunity, 106, 102308.

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Purification of VHL Isoforms

Cited 1 time

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There is conflicting nomenclature used by different investigators to assign pVHL isoforms: pVHL₂₁₃ is also known as pVHL₃₀ or pVHL₂₅, while pVHL₁₆₀ is also referred as pVHL₁₉ or pVHL₂₁. In this study, to prevent this confusion, each isoform will be named according to the number of amino acids of the molecule, that is, pVHL₂₁₃, pVHL₁₆₀ and pVHL₁₇₂.
All recombinant proteins were prepared using the E.coli strain BL21(DE3)pLysS transformed with pET21a-hVHL213, hVHL172 or hVHL160 and induced to express the proteins VHL₂₁₃(His)₆, VHL₁₇₂(His)₆ and VHL₁₆₀(His)₆ with IPTG. Proteins were prepared and purified by Talon affinity chromatography following the manufacturer's instructions (Clontech, Mountain View, CA, USA) as described by Martin et al (2013) (link). The purity of the eluted fractions was assessed by 12.5% SDS–PAGE and silver staining.

Chesnel F., Hascoet P., Gagné J.P., Couturier A., Jouan F., Poirier G.G., Le Goff C., Vigneau C., Danger Y., Verite F., Le Goff X, & Arlot-Bonnemains Y. (2015). The von Hippel–Lindau tumour suppressor gene: uncovering the expression of the pVHL172 isoform. British Journal of Cancer, 113(2), 336-344.

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Selenomethionine Labeling and Purification of Yeast Tah1

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For selenomethionine (SeMet) labelling, Escherichia coli Rosetta DE3 cells containing the appropriate expression vector were grown in minimal medium supplemented with SeMet. Synthetic full-length yeast Tah1 was expressed from pRSET-A. Tah1 was purified by Talon affinity chromatography (Clontech, Oxford, England) equilibrated in 20 mM Tris pH 7.5 containing 100 mM NaCl and eluted with the same buffer but containing 500 mM imidazole at pH 7.0. The protein was then concentrated using Vivaspin concentrators (5000 Da molecular-weight cutoff) and subjected to Superdex 75 HR gel-filtration chromatography equilibrated in 20 mM Tris pH 7.5 containing 500 mM NaCl, 0.5 mM TCEP. Pure Tah1 was dialyzed in 10 mM HEPES pH 7.5, 140 mM NaCl, 0.5 mM TCEP and then concentrated to 10 mg ml⁻¹ in the presence of 5 mM Hsp90 peptide (EDASRMEEVD) and stored frozen at −20°C. Purification of AIP was as previously described (Morgan et al., 2012 ▶ ).

Morgan R.M., Pal M., Roe S.M., Pearl L.H, & Prodromou C. (2015). Tah1 helix-swap dimerization prevents mixed Hsp90 co-chaperone complexes. Acta Crystallographica Section D: Biological Crystallography, 71(Pt 5), 1197-1206.

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