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ProBond™ Purification System
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Citations & References (57)
Invitrogen™

ProBond™ Purification System

ProBond™精製システムは、ポリヒスチジン（6xHis）配列を含む組換えタンパク質の精製キットです。 このキットはInvitrogen社のProBond™ニッケルキレートレジンを用いており、様々な条件下で精製できるネイティブおよび変性バッファーが含まれます。詳細を見る
テクニカルサポートオーダーサポート
製品番号（カタログ番号）数量
K850016 purifications
製品番号（カタログ番号） K85001
価格（JPY）
114,300
Each
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数量:
6 purifications
一括またはカスタム形式をリクエストする
ProBond™精製システムは、ポリヒスチジン（6xHis）配列を含む組換えタンパク質の精製キットです。 このキットはInvitrogen社のProBond™ニッケルキレートレジンを用いており、様々な条件下で精製できるネイティブおよび変性バッファーが含まれます。
研究用途にのみご使用ください。診断目的には使用できません。
仕様
フォーマットSuspension
製品タイプProBond™ Purification System
数量6 purifications
製品ラインProBond
タンパク質タグHis Tag
Unit SizeEach
組成および保存条件
Six 2-ml resin columns and buffers for native and denaturing purification. Twelve milliliters of ProBond™ pre-charged resin. Store at +4°C. All reagents are guaranteed stable for 6 months when properly stored.

よくあるご質問（FAQ）

How do you typically detect expression of a recombinant fusion protein?

Western blot analysis is typically used to detect the expressed protein. We sell several antibodies against various epitopes, such as Xpress, HisG, V5, or C-terminal 6xHis. Additionally, His-tagged proteins can be purified using our ProBond Purification System via affinity purification.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

After I elute my protein from the ProBond column under denaturing conditions and I dialyze it to remove the urea, my protein precipitates. Any suggestions on how to correct this?

There are a few suggestions provided from our R&D scientists:

(1) Use stepwise dialysis against successively lower concentrations of urea buffers to slow the refolding.

(2) Add glycerol to the folding buffer; usually between 10 to 50%, but the amount must be determined empirically.

(3) After the denaturing washes, do several washes under native conditions and then elute under native conditions.

Note: you may also try to rescue a precipitated protein by adding denaturing buffer and either trying the glycerol dialysis or rebinding to the column.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

What expression levels can be expected with the pTrcHis/CAT construct?

In one experiment, 35 µg CAT/mg total protein or 68.2 mg CAT/liter of culture was obtained. 50 ml of cell extract was loaded onto a ProBond column and 2 mg of CAT was recovered. The eluted protein appeared reasonably pure on a gel.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

I purified my protein from a ProBond column using denaturing conditions. After elution, I tried digesting off my N- terminal tag with EKMax Enterokinase, but see no EK cleavage. What can you suggest I try?

The enzyme could be denatured. Try buffer exchange or dialysis before digestion with EKMax Enterokinase.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Can ProBond or Ni-NTA beads be used for large-scale preparations?

ProBond and Ni-NTA beads can be used in FPLC columns. However, the beads can only withstand low pressure (~43.5 psi max).

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

View all ProBond™ Purification System FAQs

引用および参考文献 (57)

引用および参考文献
Abstract
Cellular uptake of saposin (SAP) precursor and lysosomal delivery by the low density lipoprotein receptor-related protein (LRP).
Authors:Hiesberger T, Huttler S, Rohlmann A, Schneider W, Sandhoff K, Herz J
Journal:EMBO J
PubMed ID:9707421
'Sphingolipid activator proteins SAP-A, -B, -C and -D (also called saposins) are generated by proteolytic processing from a 73 kDa precursor and function as obligatory activators of lysosomal enzymes involved in glycosphingolipid metabolism. Although the SAP precursor can be recognized by the mannose-6-phosphate (M-6-P) receptor and shuttled directly from the ... More
RasGRP4, a new mast cell-restricted Ras guanine nucleotide-releasing protein with calcium- and diacylglycerol-binding motifs. Identification of defective variants of this signaling protein in asthma, mastocytosis, and mast cell leukemia patients and demonstration of the importance of RasGRP4 in mast cell development and function.
Authors: Yang Yi; Li Lixin; Wong Guang W; Krilis Steven A; Madhusudhan M S; Sali Andrej; Stevens Richard L;
Journal:J Biol Chem
PubMed ID:11956218
'A cDNA was isolated from interleukin 3-developed, mouse bone marrow-derived mast cells (MCs) that contained an insert (designated mRasGRP4) that had not been identified in any species at the gene, mRNA, or protein level. By using a homology-based cloning approach, the approximately 2.6-kb hRasGRP4 transcript was also isolated from the ... More
Expression of a 28-kilodalton glutathione S-transferase antigen of Schistosoma mansoni on the surface of filamentous phages and evaluation of its vaccine potential.
Authors:Rao KV, He YX, Kalyanasundaram R,
Journal:Clin Diagn Lab Immunol
PubMed ID:12853382
'A cloning and expression system that allows display of proteins on the surface of filamentous phages was exploited to display a 28-kDa glutathione S-transferase (Sm28GST) antigen of the human parasite Schistosoma mansoni. The phage-displayed Sm28GST (pdGST) was immunoreactive and was recognized by immune sera, suggesting that the Sm28GST protein displayed ... More
The cathepsin B of Toxoplasma gondii, toxopain-1, is critical for parasite invasion and rhoptry protein processing.
Authors: Que Xuchu; Ngo Huân; Lawton Jeffrey; Gray Mary; Liu Qing; Engel Juan; Brinen Linda; Ghosh Partho; Joiner Keith A; Reed Sharon L;
Journal:J Biol Chem
PubMed ID:12000756
'Cysteine proteinases play a major role in invasion and intracellular survival of a number of pathogenic parasites. We cloned a single copy gene, tgcp1, from Toxoplasma gondii and refolded recombinant enzyme to yield active proteinase. Substrate specificity of the enzyme and homology modeling identified the proteinase as a cathepsin B. ... More
Proapoptotic activity of Caenorhabditis elegans CED-4 protein in Drosophila: implicated mechanisms for caspase activation.
Authors:Kanuka H, Hisahara S, Sawamoto K, Shoji S, Okano H, Miura M
Journal:Proc Natl Acad Sci U S A
PubMed ID:9874786
'CED-4 protein plays an important role in the induction of programmed cell death in Caenorhabditis elegans through the activation of caspases. However, the precise mechanisms by which it activates caspases remain unknown. To investigate the conservation of CED-4 function in evolution, transgenic Drosophila lines that express CED-4 in the compound ... More
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