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Alexa Fluor® 488 Mouse Anti-p38 MAPK (pT180/pY182)
Alexa Fluor® 488 Mouse Anti-p38 MAPK (pT180/pY182)
Flow cytometric analysis of p38 MAPK (pT180/pY182). Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (unshaded) or stimulated (shaded) with 40 nM PMA for 10 minutes at 37°C. Cells were fixed with BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37°C and then permeablized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) for 30 minutes on ice. Cells were then washed twice in BD Pharmingen™ Stain Buffer and then stained with the Alexa Fluor® 488 mouse anti-p38 MAPK (pT180/pY182) antibody. The cells were analyzed on a BD FACSCalibur™ flow cytometer. For intracellular staining of human whole blood, BD Phosflow™ Lyse/Fix buffer (Cat. No. 558049) may be used for fixation.
Flow cytometric analysis of p38 MAPK (pT180/pY182). Human peripheral blood mononuclear cells (PBMCs) were either left unstimulated (unshaded) or stimulated (shaded) with 40 nM PMA for 10 minutes at 37°C. Cells were fixed with BD Cytofix™ buffer (Cat. No. 554655) for 10 minutes at 37°C and then permeablized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) for 30 minutes on ice. Cells were then washed twice in BD Pharmingen™ Stain Buffer and then stained with the Alexa Fluor® 488 mouse anti-p38 MAPK (pT180/pY182) antibody. The cells were analyzed on a BD FACSCalibur™ flow cytometer. For intracellular staining of human whole blood, BD Phosflow™ Lyse/Fix buffer (Cat. No. 558049) may be used for fixation.
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BD Phosflow™
MAPK14; p38 MAP kinase; p38 MAPK; RK; CSBP2
Human (QC Testing), Mouse,Rat (Tested in Development)
Mouse IgG1, κ
Phosphorylated Human p38 MAPK (pT180/pY182) Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_399877
Aqueous buffered solution containing BSA 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 was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.

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For more information about Phosflow:  Please refer to http://www.bdbiosciences.com/documents/phosflow_technology_product_list.pdf

Investigators may also find the following protocols to be helpful:

Phosflow protocol for human PBMC:  Please refer to http://www.bdbiosciences.com/documents/Phosflow_Protocol_for_Human_PBMCs.pdf

Phosflow protocol for human whole blood:  Please refer to http://www.bdbiosciences.com/documents/Phosflow_Protocol_for_Human_Whole_Blood_Samples.pdf

Phosflow protocol for adherent cells:  Please refer to http://www.bdbiosciences.com/support/resources/protocols/protocol_adherent.jsp

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. 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.
  3. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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.
612594 Rev. 8
抗体の詳細
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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.

612594 Rev. 8
フォーマットの詳細
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
612594 Rev.8
引用&参考文献
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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
612594 Rev. 8

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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.