His-Tag (6-Histidine) (Biotin)

Insertion of a 6-histidine tag creates a stable fusion product that does not interfere with the bioactivity of the protein or with the biodistribution of the His-tagged product. Such proteins are purified by immobilized metal ion afftinity chromatography which makes use of its high affinity for transition metal ion.\n\nApplications: \nSuitable for use in ELISA and Western Blot. Other applications not tested.\n\nRecommended Dilutions:\nELISA: 1:500-1:1000; Tested using histidine tagged ubiquitin.\nWestern Blot: 1:1000; Tested using His-tagged recombinant protein control. H5099-16A has been shown not to crossreact with any endogenous products when using bacterial extracts. Some crossreactivity may be seen with extracts of insect or mammalian cells.\nOptimal dilutions to be determined by the researcher.\n\nRecommended Secondary Reagent:\nS7973-97B: Streptavidin (HRP)\n\nHybridoma: \nNS1 myeloma cells with spleen cells from Balb/c mice. \n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

H5099-16A

Size

100ug

Applications

ELISA, WB

Hosts

Mouse

Conjugates

Biotin

Form

Supplied as a liquid in PBS, pH 7.4, 0.09% sodium azide. Labeled with Biotin.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG1

References

1. Conrath, K., et al., J. Biol. Chem. 276(10): 7346-7350 (2001). 2. Suen, J.L. et al. (2001) Immunol. 103: 301-309. 3. Hoffmann, S.C. et al. (2007) J. Biol. Chem. 282: 22370-22375. 4. Zheng, J. et al. (2007) BMC Immunol. 8: 4. 5. Bahi, A. & Dreyer, J.L. (2008) Genes Brain Behav. 7: 244-256. 6. Wrighton, K.H. et al. (2009) J. Biol. Chem. 284: 9755-9763. 7. Diefenbacher, M. et al. (2011) J Biol Chem. 286: 25027-38. 8. Alvarez, M.M. et al. (2010) Specific recognition of influenza A/H1N1/2009 antibodies in human serum: a simple virus-free ELISA method. PLoS One. 5: e10176. 9. Bahi, A. et al. (2008) Neuropsychopharmacology. 33: 2726-34. 10. Gunnarsen, K.S. et al. (2010) BMC Biotechnol. 10: 8. 11. Hwang, H.Y. et al. (2008) Mol Immunol. 45: 2570-80.

Additional Info

Recognizes proteins containing the motif H-H-H-H-H-H and proteins containing histidine tags. Reacts against all His tagged proteins so far tested. May be used to track the purification of proteins expressed in bacteria, yeast or insect cells.