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Hitrap hp chelating columns

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The HiTrap HP Chelating columns are affinity chromatography columns designed for the purification of histidine-tagged proteins. These columns contain a matrix pre-charged with metal ions, typically Ni2+ or Co2+, which can selectively bind to the histidine-tags of recombinant proteins, allowing for their efficient capture and elution.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Anti cd3 antibody, by Agilent Technologies (1 mentions)

Spelling variants of this product

HiTrap Chelating HP column (84 citations) HiTrap chelating column (37 citations) HiTrap HP column (20 citations) HiTrap Chelating HP (17 citations) HiTrap columns (11 citations) HiTrap Chelating (6 citations) HiTrap chelating HP Ni-affinity columns (2 citations)

3 protocols using hitrap hp chelating columns

1

Purification of Recombinant SEMA3F

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A full-length, His-Myc-tagged human SEMA3F construct was transfected into 293T cells using FuGENE HD Transfection Reagent (Roche, Basel, Switzerland). SEMA3F secreted into culture medium was purified on HiTrap HP Chelating columns (GE Healthcare Bio-Sciences Corp., Pittsburgh, PA)60 (link).
Nakayama H., Bruneau S., Kochupurakkal N., Coma S., Briscoe D.M, & Klagsbrun M. (2015). Regulation of mTOR Signaling by Semaphorin 3F-Neuropilin 2 Interactions In Vitro and In Vivo. Scientific Reports, 5, 11789.
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2

Antibody and Reagent Sources for Cell Signaling

Cited 1 time
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Anti-human UNC5A and -Neogenin antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX), anti-UNC5B or -DCC from Abcam (Cambridge, MA) and anti-CD3 antibody from Dako (Carpinteria, CA). Goat anti-mouse Alexa fluor 488 and goat anti-rabbit Alexa fluor 594 were purchased from Life Technologies (Carlsbad, CA). Human Netrin-1 was produced as previously described (23 (link)). Briefly, 293T cells were transfected with netrin-1/pcDNA3.1/V5-His-TOPO plasmid using FuGENE HD Transfection Reagent (Roche Applied Science, Indianapolis, IN) and secreted Netrin-1 was purified from the culture medium using HiTrap HP Chelating columns (GE Healthcare Bio-Sciences, Pittsburgh, PA), as described (28 (link)). Anti-human CD3 and anti-human CD28 were purchased from BD Pharmingen (San Jose, CA) and Phytohaemagglutinin (PHA) from Fisher Scientific (Pittsburgh, PA).
Boneschansker L., Nakayama H., Eisenga M., Wedel J., Klagsbrun M., Irimia D, & Briscoe D.M. (2016). NETRIN-1 AUGMENTS CHEMOKINESIS IN CD4+ T CELLS IN VITRO AND ELICITS A PROINFLAMMATORY RESPONSE IN VIVO. Journal of immunology (Baltimore, Md. : 1950), 197(4), 1389-1398.
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3

Purification of Recombinant Semaphorins

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Recombinant SEMA3s were purified as in our previous reports [9 (link),17 (link)]. The full-length, His-Myc-tagged, chicken SEMA3A or human SEMA3F construct was transfected into 293T cells using FuGENE HD Transfection Reagent (Roche). Semaphorins secreted into culture medium were purified on HiTrap HP Chelating columns (GE Healthcare Bio-Sciences Corp.).
Procaccia V., Nakayama H., Shimizu A, & Klagsbrun M. (2014). Gleevec/imatinib, an ABL2 kinase inhibitor, protects tumor and endothelial cells from semaphorin-induced cytoskeleton collapse and loss of cell motility. Biochemical and biophysical research communications, 448(2), 134-138.
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