Product Pathways - Related Products

Epitope Tag Antibody Sampler Kit #9962

• pathway
• more info
• application references
• datasheet PDF
• MSDS PDF
• protocols

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Source
DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) # 2368	40 microliters	W IP IHC-P IF-IC	All		Rabbit
GST (91G1) Rabbit mAb # 2625	40 microliters	W IP IF-IC	All		Rabbit
HA-Tag (6E2) Mouse mAb # 2367	40 microliters	W IP IHC-P IF-IC F	All		Mouse
His-Tag (27E8) Mouse mAb # 2366	40 microliters	W IP	All		Mouse
Myc-Tag (71D10) Rabbit mAb # 2278	40 microliters	W IP IF-IC F	All		Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074	100 microliters				Goat
Anti-mouse IgG, HRP-linked Antibody # 7076	50 microliters				Horse

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  IC-ABC=Immunocytochemistry (ABC)  F=Flow Cytometry

Specificity / Sensitivity

All antibodies in the Epitope Tag Antibody Sampler Kit detect overexpressed fusion proteins containing the corresponding epitope tags. DYKDDDDK Tag Antibody recognizes the DYKDDDDK peptide (the same epitope recognized by Sigma's Anti-FLAG^® antibodies), and its binding specificity is NOT dependent on the presence of divalent metal cations.

Source / Purification

Mouse monoclonal antibodies are produced by immunizing mice with a 6xHis synthetic peptide (KLH-coupled) or a synthetic peptide containing the influenza hemagglutinin epitope (YPYDVPDYA) (KLH-coupled). Rabbit monoclonal antibodies are producted by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to residues 410-419 of human c-Myc (EQKLISEEDL) or with a GST fusion protein. Polyclonal antibodies are produced by immunizing rabbits with a synthetic DYKDDDDK peptide (KLH-coupled). Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Epitope tags are useful for the labeling and detection of proteins using immunoblotting, immunoprecipitation and immunostaining techniques. Due to 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. Davis, A.H. et al. (1993) Biotechnology 11, 933-936.
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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

• 2368 DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody)
• 2625 GST (91G1) Rabbit mAb
• 2367 HA-Tag (6E2) Mouse mAb
• 2366 His-Tag (27E8) Mouse mAb
• 2278 Myc-Tag (71D10) Rabbit mAb
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7076 Anti-mouse IgG, HRP-linked Antibody
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.