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Alexa Fluor® 647 Mouse anti-MAPKAPK-2 (pT334)
Alexa Fluor® 647 Mouse anti-MAPKAPK-2 (pT334)
Analyses of MAPKAPK2 (pT334) expression by Human and Mouse Cells. Human Cells Panel 1a: Flow Cytometric analysis of MAPKAPK2 (pT334) in peripheral blood lymphocytes (PBL). Whole blood cells were not stimulated (dashed line histogram) or stimulated (solid line histogram) with 400 nM Phorbol 12-Myristate 13-Acetate (PMA; Sigma, Cat. No. P8139; 15 min, 37°C). Cells were fixed in 1X BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049; 10 min, 37˚C) and permeabilized in BD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice (30 min). Cells were stained with BD Phosflow™ Alexa Fluor® 647 Mouse Anti-MAPKAPK2 (pT334) (Cat. No. 562472) Ab. Fluorescence histograms showing MAPKAPK2 (pT334) expression were generated for gated events with the light scatter characteristics of intact lymphocytes using a BD FACSCanto™ II Flow Cytometer. Panel 1b: Western blot analysis of MAPKAPK2 (pT334) expressed by peripheral blood mononuclear cells (PBMC). Lysates from 1X10^6 untreated (C) and PMA-treated (T) PBMC were blotted using Purified Mouse Anti-MAPKAPK2 (pT334) Ab (2 µg/ml, Cat. No. 562469), HRP Goat Anti-Mouse Ig (Cat. No. 554002) and a chemiluminescent detection system. MAPKAPK2 (pT334) were identified as ~49 kDa bands by Western blotting. Mouse Cells Panel 2: Splenocytes were not stimulated (dashed line histogram) or stimulated (solid line histogram) with PMA (50 nM, 15 min, 37°C). Cells were fixed, permeabilized, stained with Alexa Fluor® 647 Mouse anti-MAPKAPK2 (pT334) Ab and analyzed by flow cytometry as described above.
Analyses of MAPKAPK2 (pT334) expression by Human and Mouse Cells. Human Cells Panel 1a: Flow Cytometric analysis of MAPKAPK2 (pT334) in peripheral blood lymphocytes (PBL). Whole blood cells were not stimulated (dashed line histogram) or stimulated (solid line histogram) with 400 nM Phorbol 12-Myristate 13-Acetate (PMA; Sigma, Cat. No. P8139; 15 min, 37°C). Cells were fixed in 1X BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049; 10 min, 37˚C) and permeabilized in BD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice (30 min). Cells were stained with BD Phosflow™ Alexa Fluor® 647 Mouse Anti-MAPKAPK2 (pT334) (Cat. No. 562472) Ab. Fluorescence histograms showing MAPKAPK2 (pT334) expression were generated for gated events with the light scatter characteristics of intact lymphocytes using a BD FACSCanto™ II Flow Cytometer. Panel 1b: Western blot analysis of MAPKAPK2 (pT334) expressed by peripheral blood mononuclear cells (PBMC). Lysates from 1X10^6 untreated (C) and PMA-treated (T) PBMC were blotted using Purified Mouse Anti-MAPKAPK2 (pT334) Ab (2 µg/ml, Cat. No. 562469), HRP Goat Anti-Mouse Ig (Cat. No. 554002) and a chemiluminescent detection system. MAPKAPK2 (pT334) were identified as ~49 kDa bands by Western blotting. Mouse Cells Panel 2: Splenocytes were not stimulated (dashed line histogram) or stimulated (solid line histogram) with PMA (50 nM, 15 min, 37°C). Cells were fixed, permeabilized, stained with Alexa Fluor® 647 Mouse anti-MAPKAPK2 (pT334) Ab and analyzed by flow cytometry as described above.
商品详情
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BD Phosflow™
MAP kinase-activated protein kinase 2, MAPKAP kinase 2; MK2; MAPK2
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG1, κ
Phosphorylated Human MAPKAPK-2 Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_11154037
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


准备和存储

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

商品通知

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  6. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  7. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  9. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
562472 Rev. 1
抗体详情
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P24-694

The P24-694 monoclonal antibody specifically binds to the phosphorylated T334 site (pT334) of MAPKAPK-2. MAPKAPK-2 is a serine/threonine protein kinase. This ~49 kDa member of the MAPKAPK family of protein kinases is also known as mitogen-activated protein kinase-activated protein kinase 2. MAPKAPK-2 is phosphorylated and activated by p38 MAP kinase in response to stress, cytokines and chemokines. MAPKAPK-2 is phosphorylated on multiple sites including Thr222, Ser272 and Thr334. Phosphorylation of any two of these three amino acid residues seems to be required for the activation of this kinase that  serves multiple cellular functions. Phosphorylation of Thr334 was reported to be essential for nuclear export of the heterodimer formed between p38 MAPK and MAPKAPK-2. Mice deficient in MAPKAPK-2 have been shown to be protected from ischemic injury. MAPKAPK-2 is also reported to serve as a cell cycle checkpoint kinase in response to UV irradiation. The heat shock protein, HSP27 was shown to be one of the major substrates of MAPK and MAPKAPK-2.

562472 Rev. 1
格式详情
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
562472 Rev.1
报价单和参考
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研发参考 (8)

  1. Clifton AD, Young PR, Cohen P. A comparison of the substrate specificity of MAPKAP kinase-2 and MAPKAP kinase-3 and their activation by cytokines and cellular stress. FEBS Lett. 1996; 392(3):209-214. (Biology). 查看参考
  2. Engel K, Kotlyarov A, Gaestel M. Leptomycin B-sensitive nuclear export ofMAPKAP kinase 2 is regulated by phosphorylation. EMBO J. 1998; 17(12):3363. (Biology). 查看参考
  3. Heidenreich O, Neininger A, Schratt G, et al. MAPKAP kinase 2 phosphorylates serum response factor in vitro and in vivo. J Biol Chem. 1999; 274(20):14434-14443. (Biology). 查看参考
  4. Krump E, Sanghera JS, Pelech SL, Furuya W, Grinstein S. Chemotactic peptideN-formyl-met-leu-phe activation of p38 mitogen-activated protein kinase (MAPK)and MAPK-activated protein kinase-2 in human neutrophils. J Biol Chem. 1997; 272(2):937. (Biology). 查看参考
  5. Manke IA, Nguyen A, Lim D, Stewart MQ, Elia AE, Yaffe MB. MAPKAP kinase-2 is acell cycle checkpoint kinase that regulates the G2/M transition and S phaseprogression in response to UV irradiation. Mol Cell. 2005; 17(1):37. (Biology). 查看参考
  6. Meng W, Swenson LL, Fitzgibbon MJ, et al. Structure of mitogen-activated protein kinase-activated protein (MAPKAP) kinase 2 suggests a bifunctional switch that couples kinase activation with nuclear export. J Biol Chem. 2002; 277(40):37401-37405. (Biology). 查看参考
  7. Rouse J, Cohen P, Trigon S, et al. A novel kinase cascade triggered by stress and heat shock that stimulates MAPKAP kinase-2 and phosphorylation of the small heat shock proteins. Cell. 1994; 78(6):1027-1037. (Biology). 查看参考
  8. Wang X, Xu L, Wang H, Young PR, Gaestel M, Feuerstein GZ.. Mitogen-activatedprotein kinase-activated protein (MAPKAP) kinase 2 deficiency protects brain fromischemic injury in mice. J Biol Chem. 2002; 277(46):43968. (Biology). 查看参考
查看所有文件 (8) 查看更少内容
562472 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.