His-tag 蛋白質純化產品特輯 | Abcam

His-tag 蛋白質純化產品特輯

在使用原核細胞(如大腸桿菌)或真核細胞(如酵母菌、昆蟲細胞或哺乳動物細胞)大量表現重組蛋白質時,通常會在目標蛋白基因的末端銜接上一段特殊可辨識的標籤序列(如 His-tag 或 GST-tag),以便後續藉由親和性層析技術純化取得目標蛋白進行結構或功能分析。

His-tag 標籤通常是由 6~9 個 Histidine 胺基酸所組成,由於 His-tag 標籤短小(常用的 6x-His tag 標籤分子量約僅有 0.8 kDa)不易影響目標蛋白質的摺疊構形,且能夠非常容易地藉由螯合有二價陽離子的層析膠體純化分離出來,因此成為目前最廣為使用的重組蛋白質融合標籤之一。

His-tag fused N or C terminal of the recombinant protein.

常見的 His-tag 蛋白質純化膠體包含鎳離子 (Ni²⁺) 或鈷離子 (Co²⁺) 螯合膠體,一般來說使用鎳離子螯合膠體可以取得較多的純化蛋白質產量,鈷離子螯合膠體則可以提供更佳的純化蛋白質純度。以下就為您整理報導最受科研人員們所青睞的 His-tag 蛋白質純化實驗明星商品。

Ni-NTA Affinity Resin - Amintra

由 Expedeon (part of Abcam) 所開發的 Ni-NTA Affinity Resin - Amintra (ab270549) 能夠自細胞裂解液中快速有效地純化出融合有 His-tag 標籤的重組蛋白,其產品特點包含:

  • 高蛋白質結合力 >40 mg 6x His-tagged protein
  • 天然 (native) 或變性 (denature) 條件皆可適用
  • 純化產物適用於蛋白質結構解析或功能活性分析等實驗

SDS-PAGE and western blot analysis of 8XHis tagged RBD protein of SARS-CoV-2 produced in N. benthamiana. The crude proteins were extracted from plants, 8XHis tagged RBD protein was purified by Ni–NTA affinity resin and analyzed on SDS-PAGE gels and visualized with InstantBlue staining (a). Lane M: protein ladder; Lane 1: total soluble protein of N. benthamiana agroinfiltrated with pBY2e-SARS-CoV-2 RBD; Lane 2: purified SARS-CoV-2 RBD. For western blot analysis, proteins on the blot were probed with rabbit anti-His conjugated with HRP (b). Lane 1: crude extract from non-infiltrated N. benthamiana; Lane 2: total soluble protein of N. benthamiana agroinfiltrated with pBY2e-SARS-CoV-2 RBD; Lane 3: purified SARS-CoV-2 RBD. IMAGE © Sci Rep. 2020 Oct 19;10(1):17698. Figure S1.

產品相關文獻:

  • Tit-Oon P, et al. Prediction of the binding interface between monoclonal antibody m102.4 and Nipah attachment glycoprotein using structure-guided alanine scanning and computational docking. Sci Rep. 2020 Oct 26;10(1):18256. PMID: 33106487
  • Rattanapisit K, et al. Rapid production of SARS-CoV-2 receptor binding domain (RBD) and spike specific monoclonal antibody CR3022 in Nicotiana benthamiana. Sci Rep. 2020 Oct 19;10(1):17698. PMID: 33077899
  • Taylor RL, et al. Loss-of-Function Mutations in the CFH Gene Affecting Alternatively Encoded Factor H-like 1 Protein Cause Dominant Early-Onset Macular Drusen. Ophthalmology. 2019 Oct;126(10):1410-1421. PMID: 30905644

訂購資訊:

Product Name Cat. No. Pack Size
Ni-NTA Affinity Resin - Amintra ab270549 10 ml
25 ml
100 ml

相關產品:

Category Product Name Cat. No. Pack Size
Affinity Resin Cobalt NTA Affinity Resin - Amintra ab270018 10 ml
25 ml
100 ml
Prepacked Column His-Tag Protein Purification Column ab270528 5 x 1ml
ab270529 5 x 5ml
Antibody Anti-6X His tag® antibody [HIS.H8] ab18184 100 µg
Anti-6X His tag® antibody ab9108 100 µg
Recombinant Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 100 µl
Anti-6X His tag® antibody (HRP) ab1187 100 µg
Anti-6X His tag® antibody (FITC) ab1206 250 µg
ELISA Kit GST 6XHis-tag ELISA Kit ab128573 1 x 96 tests
Protein 6X His tag® peptide ab14943 100 µg
500 µg

BaseMuncher Endonuclease

由 Expedeon (part of Abcam) 所開發的 BaseMuncher Endonuclease (ab270049) 是蛋白質純化實驗的絕佳助手,能夠專一性水解細胞裂解液中的所有 DNA、RNA,降低樣本黏稠度,提升蛋白質純化效率!產品特點包含:

  • >99% 高純度核酸水解酶,不具蛋白水解活性
  • 活性為 DNAase 的 100 倍
  • 超高 C/P 值!具有與 Benzonase 同等的品質與效率,但價格更便宜

DNA digestion with BaseMuncher Endonuclease or nuclease was monitored by agarose gel. 50 mg of salmon sperm DNA was incubated with the indicated units of BaseMuncher Endonuclease (ab270049) or another brand of nuclease at 37°C for 30 mins in a buffer containing 50 mM Tris-HCl; pH 8.0 and 1 mM MgCl₂.

產品相關文獻:

  • Rodríguez-Martínez M, et al. Evidence That STK19 Is Not an NRAS-dependent Melanoma Driver. Cell. 2020 Jun 11;181(6):1395-1405.e11. PMID: 32531245
  • Jones M, et al. Cryo-EM structures of the XPF-ERCC1 endonuclease reveal how DNA-junction engagement disrupts an auto-inhibited conformation. Nat Commun. 2020 Feb 28;11(1):1120. PMID: 32111838
  • Sherlock D, et al. Identification of the First Gene Transfer Agent (GTA) Small Terminase in Rhodobacter capsulatus and Its Role in GTA Production and Packaging of DNA. J Virol. 2019 Nov 13;93(23):e01328-19. PMID: 31534034

訂購資訊:

Product Name Cat. No. Pack Size
BaseMuncher Endonuclease ab270049 25,000 units
100,000 units

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