Description
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.
Format
- Format
Alexa Fluor® 488
- Excitation Source
Blue 488 nm
- Excitation Max
495 nm
- Emission Max
519 nm
Alexa Fluor® conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of FITC. However, Alexa Fluor® 488 tends to be brighter and more optimal for intracellular applications. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. Alexa Fluor® 488 exhibits extraordinary photostability, which makes it highly suitable for fluorescence microscopy.
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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.
Product Notices
- 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).
- 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.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
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
