Revision 2

#9962Store at -20C

1 Kit

(5 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
GST-Tag (91G1) Rabbit mAb 2625 20 µl Rabbit IgG
Myc-Tag (71D10) Rabbit mAb 2278 20 µl Rabbit IgG
HA-Tag (C29F4) Rabbit mAb 3724 20 µl Rabbit IgG
His-Tag (D3I1O) XP® Rabbit mAb 12698 20 µl Rabbit IgG
DYKDDDDK Tag (9A3) Mouse mAb (Binds to same epitope as Sigma-Aldrich Anti-FLAG M2 antibody) 8146 20 µl Mouse IgG1
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Epitope Tag Antibody Sampler Kit provides an economical means to analyze the expression of a variety of epitope tagged proteins. The kit contains enough primary and secondary antibodies to perform two Western blots per primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation, and immunostaining techniques. Because of their small size, they are unlikely to affect the tagged protein’s biochemical properties.

Several different epitope tags are now commonly utilized and readily available. For instance, a variety of plasmids contain DNA that encodes an amino-terminal tag consisting of six histidine (6xHis) residues followed by an extended multiple cloning site. The 6xHis tag on the expressed recombinant proteins allows for efficient coupling to Ni2+ affinity resins and purification by single step chromatography (1). As is the case with other protein tag systems (2), this polyhistidine tag can often be cleaved at sites recognized by proteases such as thrombin and enterokinases to isolate the protein of interest (1). Glutathione S-transferase (GST) is another widely used fusion partner, since it provides both an easily detectable Tag and a simple purification process with little effect on the biological function of the protein of interest. Numerous vectors containing GST-Tag have been developed for both prokaryotic and eukaryotic systems over the past decade (3-5). The HA tag, derived from an epitope of the influenza hemagglutinin protein, has also been extensively used as a general epitope tag in expression vectors (6), while the Myc epitope tag is routinely used to detect expression of recombinant proteins in bacteria, yeast, insect and mammalian cell systems (7). Finally, the DYKDDDDK peptide has been used extensively as a general epitope tag in expression vectors and consists of only eight amino acids. This peptide can be expressed and detected with the protein of interested as an amino-terminal or carboxy-terminal fusion (8).

  1. Kroll, D.J. et al. (1993) DNA Cell Biol. 12, 441-453.
  2. di Guan, C. et al. (1988) Gene 67, 21-30.
  3. Guan, K.L. and Dixon, J.E. (1991) Anal. Biochem. 192, 262-267.
  4. Davies, A.H. et al. (1993) Biotechnology (N Y) 11, 933-6.
  5. Yu, J. et al. (1998) Mol. Cell. Biol. 18, 1379-1387.
  6. Field, J. et al. (1988) Mol. Cell. Biol. 8, 2159-2165.
  7. Munro, S. and Pelham, H.R. (1984) EMBO J. 3, 3087-3093.
  8. Brizzard, B. L. et al. (1994) Biotechniques 16, 730-735.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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