ProBond™ Purification System
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Citas y referencias (57)
Invitrogen™

ProBond™ Purification System

El sistema de purificación ProBond™ está diseñado para la purificación de proteínas recombinantes que contienen una secuencia de polihistidina (6xHis).Más información
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Número de catálogoCantidad
K850016 purificaciones
Número de catálogo K85001
Precio (USD)
1.506,60
Each
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Cantidad:
6 purificaciones
Pedido a granel o personalizado
Precio (USD)
1.506,60
Each
Añadir al carro de la compra
El sistema de purificación ProBond™ está diseñado para la purificación de proteínas recombinantes que contienen una secuencia de polihistidina (6xHis). Utiliza la resina quelante de níquel ProBond™ de Invitrogen y se suministra con tampones nativos y desnaturalizantes para una purificación eficaz de las proteínas recombinantes en diferentes condiciones.
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
FormatoSuspensión
Tipo de productoSistema de purificación ProBond™
Cantidad6 purificaciones
Línea de productosProBond
Etiqueta de proteínaEtiqueta His
Unit SizeEach
Contenido y almacenamiento
Seis columnas de resina de 2 ml y tampones para purificación nativa y desnaturalizante. Doce mililitros de resina precargada ProBond™. Almacenar a +4 °C. Se garantiza la estabilidad de todos los reactivos durante 6 meses si se almacenan correctamente.

Preguntas frecuentes

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

Citations & References (57)

Citations & References
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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