Technical Data
H5099-16
His-Tag (6-Histidine)
Description:
The His-tagged expression system is widely used in the production of recombinant proteins. The 6xHis tag on the expressed recombinant proteins allows for efficient coupling to Ni++ affinity resins and purification by single-step chromatography. The His-tag can be cleaved at sites recognized by enzymes such as thrombin and enterokinases to isolate the protein of interest.

Applications:
Suitable for use in ELISA, Immunohistochemistry, Immunoprecipitation and Western Blot. Suitable for tracking the purification of proteins expressed in bacteria, yeast or insect cells. Other applications not tested.

Recommended Dilutions:
Western Blot: 1:500-1:1000
Immunohistochemistry: Frozen sections
Optimal working dilutions to be determined by researcher.

Recommended Secondary Reagents:
I1904-06C: IgG, H&L (HRP) (X-Adsorbed) Pab Gt x Mo
I1904-06H: IgG, H&L (HRP) (X-Adsorbed) Pab Gt x Mo
I1904-65C: IgG, H&L (HRP) Pab Rb x Mo
I1904-65H: IgG, H&L, X-Adsorbed (HRP) Pab Rb x Mo
I1904-65N: IgG (HRP) Pab Rb x Mo
I1903-07E: IgG (HRP) Pab Rb x Mo
I1904-67R: IgG H&L, X-adsorbed (HRP) Pab Rb x Mo

Hybridoma:
NS1 myeloma cells with spleen cells from Balb/c mice.

Source:
Tissue cuture supernatant

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
MabIgG13H2201Affinity Purified
SizeStorageShippingSourceHost
250ug-20°CBlue IceMouse
Concentration:
~1mg/ml
Immunogen:
PAX6 transcription factor linked to 6-histidine tag (HHHHHH).
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a liquid in PBS, pH 7.4, 0.09% sodium azide.
Specificity:
Recognizes proteins containing the motif H-H-H-H-H-H and proteins containing histidine tags. Suitable for tracking the purification of proteins expressed in bacteria, yeast or insect cells. , Recognizes all His tagged proteins so far tested. In Western Blot of bacterial extracts, do not crossreact with any endogenous products, though some crossreactivity may be seen with extracts of insect or mammalian cells.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Conrath, K., et al., J. Biol. Chem. 276: 7346-7350 (2001). 2. Suen, J.L., et al., Immunol. 103: 301-309 (2001). 3. Hoffmann, S.C., et al., J. Biol. Chem. 282: 22,370-22,375 (2007). 4. Zheng, J., et al., BMC Immunol. 8: 4 (2007). 5. Bahi, A. & Dreyer, J.L., Genes Brain Behav. 7: 244-256 (2008). 6. Wrighton, K.H., et al., J. Biol. Chem. 284: 9755-9763 (2009). 7. Diefenbacher, M., et al., J. Biol. Chem. 286: 25,027-25,038 (2011). 8. Alvarez, M.M., et al., PLoS One 5: e10176 (2010). 9. Bahi, A., et al., Neuropsychopharmacology 33: 2726-2734 (2008). 10. Gunnarsen, K.S., et al., BMC Biotechnol. 10: 8 (2010). 11. Hwang, H.Y., et al., Mol. Immunol. 45: 2570-2580 (2008).