The ProBond™ Purification System is designed for the purification of recombinant proteins that contain a polyhistidine (6xHis) sequence. The kitRead more
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Catalog Number
Quantity
K85001
6 purifications
Catalog number K85001
Price (USD)
898.00
Each
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Quantity:
6 purifications
Request bulk or custom format
Price (USD)
898.00
Each
Add to cart
The ProBond™ Purification System is designed for the purification of recombinant proteins that contain a polyhistidine (6xHis) sequence. The kit utilizes Invitrogens ProBond™ nickel-chelating resin and is supplied with native and denaturing buffers for efficient purification of recombinant proteins under different conditions.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
FormatSuspension
Product TypeProBond™ Purification System
Quantity6 purifications
Product LineProBond™
Protein TagHis Tag
Unit SizeEach
Contents & Storage
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.
Frequently asked questions (FAQs)
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.
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.
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.
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.