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.
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UniProt ID:

#P27361, #P28482

Entrez-Gene Id:

5595, 5594

Product Includes Quantity Reactivity MW(kDa) Isotype
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb 4370 200 µl H M R Hm Mk Mi Dm Z B Dg Pg Sc 44, 42 Rabbit IgG
p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb 4695 200 µl H M R Hm Mk Mi Dm Z B Dg Pg Ce 42, 44 Rabbit IgG

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

Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

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

Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs, such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli, including mitogens, growth factors, and cytokines (1-3), and research investigators consider it an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family, as well as Mos and Tpl2/COT. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors, such as U0126 and PD98059.

  1. Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44.
  2. Baccarini, M. (2005) FEBS Lett 579, 3271-7.
  3. Meloche, S. and Pouysségur, J. (2007) Oncogene 26, 3227-39.
  4. Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
  5. Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.
  6. Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75.
  7. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
  8. Marais, R. et al. (1993) Cell 73, 381-93.
  9. Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
  10. Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PhosphoPlus is a registered 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
    #8201

    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet

    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from serum-starved HeLa cells treated with TPA (400 nM, 4 hours) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370. 虚拟泳道式图像(左图)显示一抗稀释比例在 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 2 Expand Image
    对 Jurkat 细胞提取物进行免疫沉淀分析。泳道 1 为 10% Input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb。使用 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb 进行蛋白质印迹法分析。Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 用作二抗。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 3 Expand Image
    使用 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 和 p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107,对未经处理或经 U0126 #9903(10 µM,1 小时)或 TPA #4174(200 nM,10 分钟)处理的 COS 细胞提取物进行蛋白质印迹分析。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 5 Expand Image
    使用 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb(上)或 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695(下),对经 λ 磷酸酶或 TPA #4174 处理(如图)的 293 NIH/3T3 和 C6 细胞提取物进行蛋白质印迹分析。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 6 Expand Image
    使用 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 和 α-Tubulin (11H10) Rabbit mAb #2125 对经 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) 或 SignalSilence® p44/42 MAPK (Erk1/2) siRNA (+) 转染的 Hek 293 细胞提取物进行蛋白质印迹分析。p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb 可确认 p44/42 的表达沉默,而 α-Tubulin (11H10) Rabbit mAb 则用作 p44/42 MAPK (Erk1/2) siRNA 上样和特异性的对照。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 7 Expand Image
    使用 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb,对石蜡包埋的人乳腺癌进行免疫组织化学分析。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 8 Expand Image
    使用 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析,显示其定位于细胞质和细胞核。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 9 Expand Image
    使用 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb,对未经处理(左)或经 λ 磷酸酶处理(右)的石蜡包埋的人肺癌进行免疫组织化学分析。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 10 Expand Image
    使用 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb 对石蜡包埋的人结肠癌细胞进行免疫组织化学分析。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 11 Expand Image
    使用 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb 对 SignalSlide Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Controls #8103( 经过 U0126 #9903(左图)或 TPA #4174(右图)处理的石蜡包埋的 NIH/3T3 细胞)进行免疫组织化学分析。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 12 Expand Image
    在对照肽(左)或 #1240 p44/42 MAPK (Erk1/2) Blocking Peptide (#4695 Specific)(右)存在的情况下,使用 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 13 Expand Image
    使用 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370(绿色)和 S6 Ribosomal Protein (54D2) Mouse mAb #2317(红色),对未经处理(上)或经 λ 磷酸酶处理(下)的果蝇卵泡进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 14 Expand Image
    使用 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb(绿色)对经 U0126 (MEK1/2 Inhibitor) #9903(左)或 PDGF(右)处理的 NIH/3T3 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)。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 15 Expand Image
    使用 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370(绿色)和 β-Actin (8H10D10) Mouse mAb #3700(红色),对饥饿过夜,然后用 U0126 #9903(10 uM,2 小时;左)或 PDBu (Phorbol 12,13-Dibutyrate) #12808(100 nM,15 分钟;右)处理的 HT1080 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 16 Expand Image
    使用 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对经 U0126(10 µM,2 小时;绿色)处理或经 TPA #4174 (200 nM,30 分钟;蓝色)处理的 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 17 Expand Image
    以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)作为对照,使用 p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb(实线)对 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
    PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet: Image 18 Expand Image
    使用 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) 对 3T3 细胞提取物进行免疫沉淀分析。细胞用 TPA(200 nM,15 分钟)处理。泳道 1 为 10% 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb。使用 Phosphop44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb 进行蛋白质印迹分析。使用 Mouse Anti-rabbit IgG (Light-Chain Specific) (D4W3E) mAb #45262 作为二抗。