Alexa Fluor® 647 Mouse anti-MEK1 (pS298)
Clone J114-64 (RUO)
- Brand BD Phosflow™
- Alternative Name MAP2K1, MAPKK1, MKK1, MP2K1, PPKMK1
- Vol. Per Test 20 µl
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Human (QC Testing) Mouse (Tested in Development)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Phosphorylated Human MEK1 Peptide
- Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
MEK (Map/Erk Kinase) 1 and 2 are serine/threonine kinases, also known as MAP kinase kinases (MAP2K1 and 2, MAPKK1 and 2, or MKK1 and 2). They activate the MAP (Mitogen-Activated Protein) kinases, also known as ERKs (Extracellular signal Regulated Kinases), which are critical kinases in multiple signal transduction pathways that regulate cell growth and differentiation. Activation of MEK 1 and 2 is dependent upon phosphorylation of serines 218 and/or 222 by activated MAP kinase kinase kinases (MAP3Ks), such as the Raf isoforms. Hormones, growth and differentiating factors, or tumor promoters induce Raf activation via activation of Ras proteins. Alternatively, cellular adhesion can lead to phosphorylation of MEK1 at serine 298 (S298), mediated by p21-activated kinase (PAK). The S298-phosphorylated MEK1 has an enhanced capacity to interact with Raf, resulting in MEK1 activation.
The J114-64 monoclonal antibody recognizes the phosphorylated S298 of MEK1.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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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® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
- 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).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Species testing during development may have been performed with a different format of the same clone. Selected applications have been tested for cross-reactivity.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Either BD Cytofix™ fixation buffer or BD Phosflow™ Fix Buffer I may be used for cell fixation.