His tag purification resin

Manufactured by Roche

Sourced in China, United States, Switzerland, Germany

His-Tag Purification Resin is a chromatography media designed for the purification of recombinant proteins containing histidine-tag (His-tag) sequences. This resin is composed of a porous agarose matrix with immobilized nickel ions (Ni2+) that selectively bind to the His-tag, allowing the target protein to be captured and separated from other components in the sample.

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Lab products found in correlation

Glutathione sepharose 4b, by GE Healthcare (2 mentions) Enzchek phospholipase a2 activity kit, by Thermo Fisher Scientific (2 mentions) Hiload 16 60 superdex 200 column, by GE Healthcare (1 mentions) Pbe 5a, by Takara Bio (1 mentions) Pbe 5u, by Takara Bio (1 mentions) Pbe 5c, by Takara Bio (1 mentions) Pbe 5g, by Takara Bio (1 mentions) Pbe 1g, by Takara Bio (1 mentions) Hitrap q hp, by GE Healthcare (1 mentions) Superose 6 increase 10 300 gl size exclusion chromatography, by Cytiva (1 mentions) Bml pw8810, by Enzo Life Sciences (1 mentions) Affinity column, by Qiagen (1 mentions) Amicon ultra 15, by Merck Group (1 mentions) Pet15b vector, by Merck Group (1 mentions) Escherichia coli bl21 gold de3, by Agilent Technologies (1 mentions) Amicon ultra device, by Merck Group (1 mentions) Superdex 75, by GE Healthcare (1 mentions) Superdex 200, by Cytiva (1 mentions)

Close spelling variants of this product

COmplete His-Tag Purification Resin His-tag

13 protocols using his tag purification resin

Purification and Characterization of PUF-8 Proteins

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All the wild-type and mutant proteins of PUF-8^172–535 and PUF-8^172–522 were expressed in Escherichia coli BL21-GOLD (DE3) cells (Novagen). Cells were grown in Luria-Bertani (LB) medium at 37°C to an OD₆₀₀ of 0.7–1.0 and induced with 0.4 mM isopropyl β-d-thiogalactopyranoside (IPTG) for 22 h at 16°C. The cells were pelleted and resuspended in lysis buffer (25 mM Tris–HCl, pH 8.5, 500 mM NaCl, 20 mM imidazole, and 5% (v/v) glycerol) and lysed by sonication. The supernatant were purified by His tag purification resin (Roche), followed by size exclusion chromatography on a HiLoad Superdex 200 16/60 column (GE Healthcare) in buffer A (25 mM Tris–HCl, pH 8.5, 250 mM NaCl).
Native RNA: PBE-5A (5′-UGUAAAUA-3′), PBE-5U (5′-UGUAUAUA-3′), PBE-5C (5′-UGUACAUA-3′), PBE-5G (5′-UGUAGAUA-3′), PBE-1G (5′-GGUAAAUA-3′), PBE-2U (5′-UUUAUAUA-3′), RNA_ctrl (5′-AUAAAUGU-3′), and the 5′-Fam-tagged RNAs (PBE-5A, PBE-5U, PBE-5C, PBE-5G, PBE-1G and Ctrl) were purchased from Takara Bio Inc. (Dalian, China). RNA oligomers were dissolved in diethyl pyrocarbonate (DEPC)-treated water to a concentration of 2 mM as the stock solution.

Xu Z., Zhao J., Hong M., Zeng C., Guang S, & Shi Y. (2021). Structural recognition of the mRNA 3′ UTR by PUF-8 restricts the lifespan of C. elegans. Nucleic Acids Research, 49(17), 10082-10097.

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Recombinant Angiogenin and RNase 4 Production

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The Angiogenin (Ang) and RNase 4 cDNA were obtained by RT-PCR from mouse liver total RNA with primers listed in Supplementary Data 1, and inserted into pET28a at the sites of Nde I and Xho I to obtain pET28a-Ang and pET28a-RNase 4. Recombinant Ang and RNase 4 were obtained by IPTG-inducible expression in Escherichia coli BL21 Rosetta (DE3) cells and purification with His-Tag Purification Resin (Roche) and ion-exchange column (Hitrap Q HP, GE Healthcare). Purified Ang and RNase 4 were analyzed by SDS-PAGE and Coomassie blue staining, and then dialyzed to storage buffer (20 mM Tris–HCl pH 7.5, 200 mM NaCl and 5% glycerol) and stored at −80 °C.

Lai H., Feng N, & Zhai Q. (2023). Discovery of the major 15–30 nt mammalian small RNAs, their biogenesis and function. Nature Communications, 14, 5796.

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Purification of SARS-CoV-2 XBB.1 Spike Ectodomain

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Protein expression and purification of XBB.1 S protein ectodomain and human ACE2 were performed as previously described^{10 (link)}. Briefly, the expression plasmid, pHLsec, encoding the XBB.1 S protein ectodomain bearing six proline substitutions (F817P, A892P, A899P, A942P, K986P and V987P)^{83 (link)} and the deletion of the furin cleavage site (i.e., RRAR to GSAG substitution) with a T4-foldon domain or soluble human ACE2 ectodomain were transfected into HEK293S GnTI(-) cells. Expressed proteins in the cell-culture supernatant were purified with a cOmplete His-Tag Purification Resin (Roche, Cat# 5893682001) affinity column, followed by Superose 6 Increase 10/300 GL size-exclusion chromatography (Cytiva, Cat# 29091596) with calcium- and magnesium-free PBS buffer.

Tamura T., Ito J., Uriu K., Zahradnik J., Kida I., Anraku Y., Nasser H., Shofa M., Oda Y., Lytras S., Nao N., Itakura Y., Deguchi S., Suzuki R., Wang L., Begum M.M., Kita S., Yajima H., Sasaki J., Sasaki-Tabata K., Shimizu R., Tsuda M., Kosugi Y., Fujita S., Pan L., Sauter D., Yoshimatsu K., Suzuki S., Asakura H., Nagashima M., Sadamasu K., Yoshimura K., Yamamoto Y., Nagamoto T., Schreiber G., Maenaka K., Hashiguchi T., Ikeda T., Fukuhara T., Saito A., Tanaka S., Matsuno K., Takayama K, & Sato K. (2023). Virological characteristics of the SARS-CoV-2 XBB variant derived from recombination of two Omicron subvariants. Nature Communications, 14, 2800.

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Purification of GST-tagged and His-tagged Proteins

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GST-tagged proteins, His-SIN3A and His-hMOF were induced using IPTG in BL21 bacterial cells and were further purified following the manual of Glutathione-Sepharose 4B (GE Health, USA) and His-tag Purification Resin (Roche, USA). GST or GST-fusion protein was incubated with the whole cell lysates or His-hMOF as indicated in results and prepared glutathione-sepharose beads at 4°C for overnight. After centrifugation at 1,500 × g for 10 min, the pellets were washed for four times with 100 bead volumes of NETN buffer (0.5% NP-40, 0.1mM EDTA, 20 mM Tris–HCl, pH 7.4, 300 mM NaCl). The pellets were eluted by heating at 100°C for 10 min in SDS-PAGE loading buffer.

Zhong J., Li X., Cai W., Wang Y., Dong S., Yang J., Zhang J., Wu N., Li Y., Mao F., Zeng C., Wu J., Xu X, & Sun Z.S. (2016). TET1 modulates H4K16 acetylation by controlling auto-acetylation of hMOF to affect gene regulation and DNA repair function. Nucleic Acids Research, 45(2), 672-684.

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Purification of Engineered Fluorescent Protein

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KatushkaS158A (tfRFP) were purified as described 28 (link). Briefly, E. coli BL21 (DE3) cells were transformed with pRSET-tfRFP. After full expression, the collected bacteria were frozen at -80 ℃ for at least 30 mins, then thawed and broken by ultrasound. tfRFP was purified by His-Tag Purification Resin (Roche, Basel, Switzerland). The obtained protein was dialyzed in PBS and diluted to a concentration of 2 mg/mL.

Dai B., Zhang R., Qi S., Liu L., Zhang X., Deng D., Zhang J., Xu Y., Liu F., Liu Z., Luo Q, & Zhang Z. (2022). Intravital molecular imaging reveals that ROS-caspase-3-GSDME-induced cell punching enhances humoral immunotherapy targeting intracellular tumor antigens. Theranostics, 12(17), 7603-7623.

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Ubiquitination Pulldown from Rapamycin-Induced Aggregates

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HEK293T cells were transfected with the mCherry-EGFP-PIM construct and pMT107 (His-Ubi) using Fugene6 according to the manufacturer’s protocol. PIM construct in GW1 vector with CMV promoter was used in these experiments for higher expression levels (Addgene #111759). One day after transfection, rapalog2 was added for 1 h. Four hour after aggregate formation, pulldown protocol was performed. Briefly, cells were washed in cold PBS after which cells were gently scraped in cold PBS with 10 mM NEM. Cells were pelleted by centrifugation and resuspended in pH 8.0 buffer (6 M guanidine HCl, 0.1 M NaH₂PO₄-Na₂HPO₄, 10 mM Tris-HCl pH 8.0, 25 mM DTT). After short sonication, input samples were collected after which cell lysates were incubated with cOmplete His-Tag purification Resin (Roche) in pH 8.0 buffer with 5 mM Imidazol at 4 °C overnight. The next day, resin was washed with pH 8.0 buffer with 0.05% Tween-20 and buffer pH 6.3 (8 M Urea, 0.1 M NaH₂PO₄-Na₂HPO₄, 10 mM Tris-HCl pH 6.3, 0.1% Tween-20, and 25 mM DTT). After the last wash, resin was pelleted and resuspended in sample buffer and boiled. Samples were loaded onto SDS-PAGE gels. Immunoblotting was performed using rabbit anti-RFP (Rockland, #600-401-379, 1/2000) and mouse anti-ubiquitin (Enzo, #BML-PW8810, 1/2000) antibodies using standard immunoblotting protocol.

Janssen A.F., Katrukha E.A., van Straaten W., Verlhac P., Reggiori F, & Kapitein L.C. (2018). Probing aggrephagy using chemically-induced protein aggregates. Nature Communications, 9, 4245.

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Producing Anti-CD3/CD20 Bispecific Antibody

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The anti-CD3/CD20 expression cassette was inserted into the empty plasmid pMC.BESXP, generating the MC-producing parental plasmid pMC.anti-CD3/CD20. Escherichia coli strain ZYCY10P3S2T, transformed with a parental plasmid, was incubated in 200 mL Terrific Broth (TB) medium at 37°C with shaking at 250 rpm for 16 h. After adding induction medium (200 mL of Luria Bertani (LB) medium, 200 μL of 20% L-arabinose, and 8.0 mL of 1 N NaOH) and incubation at 32°C with shaking at 250 rpm for 5 h, MC.CD20 was isolated using a commercially available affinity column (QIAGEN, Hilden, Germany). Purified BsAb.CD20 was a contract product made by Synbio Technologies (Suzhou, China). Briefly, Expi293F cells were transiently transfected with MC.CD20 and cultured in Expi239 expression medium (Gibco). BsAb.CD20 was purified with His Tag purification resin (Roche).

Pang X., Chen G., Huang P., Zhang P., Liu J., Hou X., He C.Y., Chen P., Xie Y.W., Zhao J, & Chen Z.Y. (2022). Anticancer effects of a single intramuscular dose of a minicircle DNA vector expressing anti-CD3/CD20 in a xenograft mouse model. Molecular Therapy Oncolytics, 24, 788-798.

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Cab45 Protein Expression and Purification

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His-Sumo-tagged Cab45 was expressed from the pI-secSUMOstar plasmid in SF9 cells and purified from cell supernatants by nickel-based affinity chromatography using an NaP, pH 8.0, buffer containing 500 mM NaCl and cOmplete His-tag Purification Resin (Roche). After elution with 250 mM imidazole, proteins were dialyzed against 20 mM Pipes, pH 6.8, containing 500 mM NaCl and 10% glycerol vol/vol for storage.

Crevenna A.H., Blank B., Maiser A., Emin D., Prescher J., Beck G., Kienzle C., Bartnik K., Habermann B., Pakdel M., Leonhardt H., Lamb D.C, & von Blume J. (2016). Secretory cargo sorting by Ca2+-dependent Cab45 oligomerization at the trans-Golgi network. The Journal of Cell Biology, 213(3), 305-314.

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Recombinant Hph5 Mutant Expression and Purification

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For protein expression,
overnight cultures of E. coli BL21
(DE3) cells harboring a pET28b-based vector were diluted into 600
mL of LB-Lennox medium and grown at 37 °C until an OD₆₀₀ of 0.6 was reached. The cytoplasmic expression of Hph5 mutants was
induced by 0.4 mM isopropyl β-d-thiogalactoside (IPTG).
The culture was grown further at 16 °C for ∼16 h with
shaking (220 rpm). After centrifugation at 4 °C for 30 min, the
cell pellet was resuspended in chilled lysis buffer (20 mM Tris, pH
7.4, 150 mM NaCl, 5 mM MgCl₂), supplemented with a 1×
protein inhibitor cocktail (Sigma).
For protein purification,
the cells were mechanically disrupted at 700 bar and the lysate was
cleared through centrifugation (10,000g) at 4 °C
for 1 h. The supernatant was incubated with 1.5 mL of His-Tag purification
resin (Roche) at 4 °C for 2 h with gentle rotation. The resin
was washed with chilled lysis buffer and resuspended in 10 mL of lysis
buffer. To remove the 6His-SUMO tag, protease ULP1 was added for on-resin
digestion, and the mixture was incubated at 4 °C for 4 h with
gentle rotation. Hph5 mutants were collected in the flow-through,
concentrated by diafiltration (Amicon Ultra-15, Millipore) and stored
at −80 °C. Protein purity and concentration were determined
by SDS-PAGE analysis and absorption at 280 nm, respectively.

Xue Z, & Quan S. (2023). Understanding the Stabilization Mechanism of a Thermostable Mutant of Hygromycin B Phosphotransferase by Protein Sector-Guided Dynamic Analysis. ACS Omega, 8(29), 25739-25748.

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Purification of GST and SIRT1 Proteins

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GST-tagged proteins, His-SIN3A and His-SIRT1, were induced using IPTG in BL21 bacterial cells and were further purified following the manual of Glutathione-Sepharose 4B (GE Health, Boston, MA, USA, Cat# 17-0756-01) and His-tag Purification Resin (Roche, Germany, Cat# 08778850001). GST or GST-fusion protein was incubated with the whole cell lysates or His-SIRT1 as indicated in the results and prepared with Glutathione-Sepharose beads at 4°C overnight. After centrifuging at 1500 g for 10 min, the pellets were washed four times with 100 bed volumes of NETN buffer (0.5% NP-40, 0.1 mM EDTA, 20 mM Tris-HCl, pH 7.4, 300 mM NaCl). The pellets were eluted by heating at 100°C for 10 min in SDS-PAGE loading buffer.

Zhang C., Zhong T., Li Y., Li X., Yuan X., Liu L., Wu W., Wu J., Wu Y., Liang R., Xie X., Kang C., Liu Y., Lai Z., Xiao J., Tang Z., Jin R., Wang Y., Xiao Y., Zhang J., Li J., Liu Q., Sun Z, & Zhong J. (2021). The hepatic AMPK-TET1-SIRT1 axis regulates glucose homeostasis. eLife, 10, e70672.

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