Revision 1
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
DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) (Alexa Fluor® 488 Conjugate) 5407 40 µl Rabbit 
GST-Tag (26H1) Mouse mAb (Alexa Fluor® 488 Conjugate) 3368 40 µl Mouse IgG2a
HA-Tag (6E2) Mouse mAb (Alexa Fluor® 488 Conjugate) 2350 40 µl Mouse IgG1
Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 488 Conjugate) 2279 40 µl Mouse IgG2a kappa
Mouse (MOPC-21) mAb IgG1 Isotype Control (Alexa Fluor® 488 Conjugate) 4878 40 µl Mouse IgG1
Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) 2975 40 µl Rabbit IgG

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

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.

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 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).

  1. Field, J. et al. (1988) Mol Cell Biol 8, 2159-65.
  2. Guan, K.L. and Dixon, J.E. (1991) Anal Biochem 192, 262-7.
  3. Davies, A.H. et al. (1993) Biotechnology (N Y) 11, 933-6.
  4. Yu, J. et al. (1998) Mol Cell Biol 18, 1379-87.
  5. Munro, S. and Pelham, H.R. (1984) EMBO J 3, 3087-93.
  6. Brizzard, B.L. et al. (1994) Biotechniques 16, 730-5.

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.

    使用に関する制限

    法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    Revision 1
    #12427

    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit

    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 1 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 2 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 3 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 4 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 5 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 6 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 7 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 8 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 9 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 10 Expand Image
    Flow cytometric analysis of COS cells, untransfected (blue) or transfected with GST (green), using GST-Tag (26H1) Mouse mAb (Alexa Fluor® 488 Conjugate.)
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 11 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 12 Expand Image
    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).
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 13 Expand Image
    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).