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BD Phosflow™ Pacific Blue™ Mouse anti-p38 MAPK (pT180/pY182)

Clone 36/p38 (pT180/pY182) (RUO)

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p38_TDS.png

Analysis of p38 MAPK (pT180/pY182) in human peripheral blood monocytes. Human peripheral blood mononuclear cells (PBMC) were either left unstimulated (open histogram) or stimulated with 40 µM Anisomycin (Calbiochem, Cat. No. 176880) for 20 minutes (shaded histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with Pacific Blue™ Mouse anti-p38 MAPK (pT180/pY182).  For data analysis, monocytes were selected by scatter profile.  Flow cytometry was performed on a BD FACSCanto™ II flow cytometer.

p38_TDS.png p38_chart2.png p38_chart2.png Blue-Cap-Violet-Label.png

Analysis of p38 MAPK (pT180/pY182) in human peripheral blood monocytes. Human peripheral blood mononuclear cells (PBMC) were either left unstimulated (open histogram) or stimulated with 40 µM Anisomycin (Calbiochem, Cat. No. 176880) for 20 minutes (shaded histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with Pacific Blue™ Mouse anti-p38 MAPK (pT180/pY182).  For data analysis, monocytes were selected by scatter profile.  Flow cytometry was performed on a BD FACSCanto™ II flow cytometer.

Product Details
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BD Phosflow™
MAPK14; p38 MAP kinase; p38 MAPK; RK; CSBP2
Human (QC Testing), Human, Mouse, Rat (Tested in Development)
Mouse IgG1, κ
Phosphorylated Human p38 MAPK (pT180/pY182) Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_1645295
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody is conjugated to Pacific Blue™ under optimum conditions, and unreacted Pacific Blue™ was removed.
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Recommended Assay Procedures

This antibody conjugate is suitable for intracellular staining of human peripheral blood mononuclear cells (using BD Cytofix™ Fixation Buffer).  Any of the three BD Phosflow™ permeabilization buffers may be used.

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Product Notices

  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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Pacific Blue™ has a maximum absorption of 416 nm and maximum emission of 451 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. 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.
  6. 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.
  7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  8. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  9. All other brands are trademarks of their respective owners.
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560313 Rev. 3
Antibody Details
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36/p38 (pT180/pY182)

Activation of the immune and inflammatory responses often involves the recognition of bacterial endotoxin (lipopolysaccharide or LPS).  Binding of LPS by monocytes results in the production and release of proinflammatory cytokines, such as IL-1 and TNF.  LPS-induced signaling cascades involve members of the Ser/Thr protein kinase family known as the Mitogen Activated Protein Kinases (MAPKs).  MAPK signal transduction pathways mediate the effects of various extracellular stimuli on biological processes such as proliferation, differentiation, and death.  The p38 MAPKs include p38α (MAPK14), β (MAPK11), γ (MAPK12), and δ (MAPK13).  These Ser/Thr kinases are activated by dual phosphorylation on threonine (T) and tyrosine (Y) within the motif Thr-Gly-Tyr located in kinase subdomain VIII.  Activation of p38 MAPK is mediated specifically by the MAP Kinase Kinases, MKK3, MKK4, and MKK6.  This leads to the activation of multiple transcription factors (NF-κB, ATF-2, Elk-1, and CHOP) that induce expression of many different genes, including proinflammatory cytokine genes.  Thus, p38 MAPKs are central kinases in multiple signal transduction pathways.

The 36/p38 (pT180/pY182) monoclonal antibody recognizes the conserved dual phosphorylated site pT180/pY182 of p38α, β, γ, and δ.

560313 Rev. 3
Format Details
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Pacific Blue
Pacific Blue™ is part of the BD violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 404-nm and an emission maximum (Em Max) at 455-nm. Pacific Blue is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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Pacific Blue
404 nm
455 nm
560313 Rev.3
Citations & References
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View product citations for antibody "560313" on CiteAb

Development References (3)

  1. Brunet A, Pouyssegur J. Identification of MAP kinase domains by redirecting stress signals into growth factor responses. Science. 1996; 272(5268):1652-1655. (Biology). View Reference
  2. Han J, Lee JD, Bibbs L, Ulevitch RJ. A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells. Science. 1994; 265(5173):808-811. (Biology). View Reference
  3. Winston BW, Chan ED, Johnson GL, Riches DW. Activation of p38mapk, MKK3, and MKK4 by TNF-alpha in mouse bone marrow-derived macrophages. J Immunol. 1997; 159(9):4491-4497. (Biology). View Reference
560313 Rev. 3

Please refer to Support Documents for Quality Certificates

 

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

 

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

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