Description
The N9-66 monoclonal antibody specifically binds to JNK1 and JNK2 phosphorylated at the pT183/pY185 sites. c-Jun NH2-terminal Kinases (JNKs), also called Stress Activated Protein Kinases (SAPKs), are mitogen-activated protein kinases (MAPKs) with observed molecular weights of ~46 kDa (JNK1) and ~54 kDa (JNK2). Along with the p38 and ERK families, JNK represents one of three major classes of MAPKs. Complete activation of JNK requires the phosphorylation of both Thr183 and Tyr185 that are located in a Thr-X-Tyr motif. Phosphorylation of these residues is carried out by MKK4 and MKK7 that are phosphorylated and activated by MEKKs and MLKs in response to stress signals delivered through small GTPases of the Rho family. Once activated, JNK can translocate into the nucleus and regulate the expression of genes through phosphorylation of c-Jun, ATF-2, and other transcription factors. JNK plays a role in signal transduction in response to cytokines and various forms of environmental stress, such as endotoxins, UV irradiation, heat, and hyperosmolarity. JNK is critical to the regulation of cell growth, apoptosis, and the cellular response to stress, making it an important factor in tumorigenesis and adaptive immunity. During antibody development, the N9-66 monoclonal antibody was found to detect phosphorylated JNK1/2 by Western blot analysis of cellular lysates and by immunofluorescent staining and flow cytometric analysis of fixed and permeabilized cells. This antibody crossreacts with phosphorylated JNK1/2 expressed by mouse cells, as tested by Western blot analysis and flow cytometry.
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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. 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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
9. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
