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Cell Signaling Technology

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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.

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    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
    使用 Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 488 Conjugate)(绿色)对 COS 亲本细胞(左图)和表达带有 Myc 标签的蛋白的 COS 细胞(右图)进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 2 Expand Image
    使用 HA-Tag (6E2) Mouse mAb (Alexa Fluor® 488 Conjugate) #2350 对转染空载(左图)或转染带有 HA 标签蛋白(右图)的 COS 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。蓝色伪彩 = DRAQ5 (DNA 荧光染料)。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 3 Expand Image
    以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate(绿色,右图)作为对照,使用 GAPDH (14C10) Rabbit mAb (Alexa Fluor® 488 Conjugate) #3906(绿色,左图)对 Cos 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 4 Expand Image
    使用 GST-Tag (26H1) Mouse mAb (Alexa Fluor® 488 Conjugate)(绿色)对转染空载(左图)或转染带有 GST 标签的蛋白(右图)的 COS 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。蓝色伪彩 = DRAQ5®(DNA 荧光染料)。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 5 Expand Image
    以 Akt (5G3) Mouse mAb (Alexa Fluor® 488 Conjugate) #2917(绿色)作为对照,使用 Mouse (MOPC-21) mAb IgG1 Isotype control (Alexa Fluor® 488 Conjugate)(红色)对未经处理的 Jurkat 细胞进行流式细胞分析。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 6 Expand Image
    使用 DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) (Alexa Fluor® 488 Conjugate)(绿色)对瞬时转染带有 DYKDDDDK 标签融合蛋白的 COS 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 7 Expand Image
    使用 Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 488 Conjugate) 对未转染(蓝色)或转染带有 Myc 标签的 Akt(绿色)的 COS 细胞进行流式细胞分析。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 8 Expand Image
    以浓度匹配的 Mouse (MOPC-21) mAb IgG1 Isotype Control (Alexa Fluor® 488 Conjugate) #4878(虚线)作为对照,使用 HA-Tag (6E2) Mouse mAb (Alexa Fluor® 488 Conjugate)(实线)对转染空载(蓝色)或转染 HA-AKT(绿色)的 293t 细胞进行流式细胞分析。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 9 Expand Image
    以未经处理(蓝色)或已经 IFN-α(绿色)处理的 Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4803 作为对照,使用 Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate)(红色)对 Jurkat 细胞进行流式细胞分析。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 10 Expand Image
    使用 GST-Tag (26H1) Mouse mAb (Alexa Fluor® 488 Conjugate) 对未转染(蓝色)或转染 GST(绿色)的 COS 细胞进行流式细胞分析。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 11 Expand Image
    以经U0126(蓝色)或 PMA(绿色)处理的 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) Mouse mAb (Alexa Fluor® 488 Conjugate) #4374 作为对照,使用 Mouse (MOPC-21) mAb IgG1 Isotype Control (Alexa Fluor® 488 Conjugate)(红色)对经过U0126处理的 Jurkat 细胞进行流式细胞分析。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 12 Expand Image
    使用 DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) (Alexa Fluor® 488 Conjugate)(绿色)对未转染(蓝色)或转染带有 DYKDDDDK 标签融合蛋白的 COS 细胞进行流式细胞分析。
    Epitope Tag Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 13 Expand Image
    以未经处理(绿色)或已经 LY294002 和 wortmannin(蓝色)处理的 Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4071 作为对照,使用 Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate)(红色)对 Jurkat 细胞进行流式细胞分析。