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12427
Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit
Antibody Conjugates
Antibody Sampler Kit

Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit #12427

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Confocal immunofluorescent analysis of parental COS cells (left) and COS cells expressing a Myc-tagged protein (right), using Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 488 Conjugate) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of COS cells, mock-transfected (left) or transfected with HA-tagged protein (right), using HA-Tag (6E2) Mouse mAb (Alexa Fluor® 488 Conjugate) #2350. Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5 (fluorescent DNA dye).
Confocal immunofluorescent analysis of Cos cells using GAPDH (14C10) Rabbit mAb (Alexa Fluor® 488 Conjugate) #3906 (green, left) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate (green, right). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent analysis of COS cells, mock transfected (left) or transfected with a GST-tagged protein (right), using GST-Tag (26H1) Mouse mAb (Alexa Fluor® 488 Conjugate) (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
Flow cytometric analysis of untreated Jurkat cells using Mouse (MOPC-21) mAb IgG1 Isotype control (Alexa Fluor® 488 Conjugate) (red) compared to Akt (5G3) Mouse mAb (Alexa Fluor® 488 Conjugate) #2917(green).
Confocal immunofluorescent analysis of COS cells transiently transfected with a DYKDDDDK-tagged fusion protein using DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) (Alexa Fluor® 488 Conjugate) (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of COS cells, untransfected (blue) or transfected with Myc-tagged Akt (green), using Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 488 Conjugate).
Flow cytometric analysis of 293t cells mock-transfected (blue) or transfected with HA-AKT (green), using HA-Tag (6E2) Mouse mAb (Alexa Fluor® 488 Conjugate) (solid lines) compared to a concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (Alexa Fluor® 488 Conjugate) #4878 (dashed lines).
Flow cytometric analysis of Jurkat cells, using Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) (red) compared to Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4803, untreated (blue) or IFN-α-treated (green).
Flow cytometric analysis of COS cells, untransfected (blue) or transfected with GST (green), using GST-Tag (26H1) Mouse mAb (Alexa Fluor® 488 Conjugate.)
Flow cytometric analysis of U0126-treated Jurkat cells using Mouse (MOPC-21) mAb IgG1 Isotype Control (Alexa Fluor® 488 Conjugate) (red) compared to Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) Mouse mAb (Alexa Fluor® 488 Conjugate) #4374, U0126-treated (blue) or PMA-treated (green).
Flow cytometric analysis of COS cells, either untransfected (blue) or transfected with a DYKDDDDK-tagged fusion protein, using DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) (Alexa Fluor® 488 Conjugate) (green).
Flow cytometric analysis of Jurkat cells, using Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) (red) compared to Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4071, untreated (green) or LY294002- and wortmannin-treated (blue).
Inquiry Info.# 12427

Kit Usage Information

Protocols

Product Description

The Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit provides an economical means to monitor the presence of proteins labeled with HA, GST, Myc, or DYKDDDDK (FLAG) tags without the need for a fluorescent secondary antibody.

Specificity / Sensitivity

GST (26H1) Mouse mAb (HRP Conjugate) recognizes transfected glutathione S-transferase (GST) fusion proteins. HA-Tag (6E2) Mouse mAb (HRP Conjugate) recognizes proteins containing the HA epitope tag. The antibody recognizes the HA-tag fused to either the amino or carboxy terminus of targeted proteins in transfected mammalian cells. Myc-Tag (9B11) Mouse mAb (HRP Conjugate) recognizes transfected proteins containing the Myc epitope tag. The antibody recognizes the Myc-tag fused to either the amino or carboxy terminus of targeted proteins in transfected mammalian cells. His-Tag (D3I1O) XP® Rabbit mAb (HRP Conjugate) recognizes recombinant proteins containing the 6xHis epitope tag. The antibody recognizes the 6xHis-tag fused to either the amino or carboxy terminus of targeted proteins in transfected cells. S-Tag (D2K2V) XP® Rabbit mAb (HRP Conjugate) recognizes transfected levels of total S-Tag protein

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide containing the influenza hemagglutinin epitope (YPYDVPDYA), a synthetic peptide corresponding to residues 410-419 of human c-Myc (EQKLISEEDL), or a GST fusion protein.

Polyclonal antibodies are produced by immunizing animals with a synthetic DYKDDDDK peptide. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

Epitope tags are useful for the labeling and detection of proteins using flow cytometry and immunostaining techniques. Because of their small size, they are unlikely to affect the tagged protein's biochemical properties. HA tag is derived from an epitope of the influenza hemagglutinin protein, which has been extensively used as a general epitope tag in expression vectors (1). Glutathione S-transferase (GST) is a 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 (2-4). Myc epitope tag is widely used to detect expression of recombinant proteins in bacteria, yeast, insect, and mammalian cell systems (5). DYKDDDDK peptide has been used extensively as a general epitope tag in expression vectors. This peptide can be expressed and detected with the protein of interest as an amino-terminal or carboxy-terminal fusion (6).

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not for Use in Diagnostic Procedures.
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