Description
MEK1 (MapK/ERK Kinase 1) is a 45-kDa member of the MEK family of dual specificity kinases.MEK is activated by a variety of cellular serine/threonine kinases including c-Raf, A-Raf, c-mos, and MEK Kinase-1.Activated MEK phosphorylates MAP kinase (ERK) at threonine and tyrosine residues.This results in activation of ERK and its signaling pathway. MEK is highly specific for ERK and various MEKs preferentially phosphorylate individual ERK isoforms.MEK1 only activates ERK1 and ERK2.This specificity may result from variations in ERK regions that are known as the phosphorylation lip and kinase backbone.MEK"s localization is cytoplasmic, but mitogenic stimulation induces a mass translocation to the nucleus.Mechanisms behind this nuclear translocation remain unknown.However, MEK contains an N-terminal nuclear export signal (NES) that mediates its rapid exodus from the nucleus and restores its unstimulated cellular distribution.
The 25/MEK1 monoclonal antibody recognizes MEK1, regardless of phosphorylation status.
The specificity of this antibody conjugate for flow cytometric analysis was validated by confirming that RNA-mediated interference (RNAi) of the specific protein reduced the staining of the cells (see figure). Furthermore, the capacity of the RNAi to down-regulate the expression of the relevant protein was confirmed by western blot analysis.
Format
- FormatAlexa Fluor® 647
- Excitation SourceRed 633 nm
- Excitation Max650nm
- Emission Max668nm
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.
Other Formats→
- PE
- Purified
Suggested Companion Products
Fixation Buffer RUO 100mLCat No: 554655
Fix Buffer I RUO 250mLCat No: 557870
Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control RUO 50TestsCat No: 558053
Perm Buffer III RUO 125mLCat No: 558050
Perm Buffer II RUO 125mLCat No: 558052
Perm/Wash Buffer I RUO 125mLCat No: 557885
Stain Buffer (FBS) RUO 500mLCat No: 554656
Stain Buffer (BSA) RUO 500mLCat No: 554657
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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.
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).
- 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.
- 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.
This mAb was characterized by flow cytometry (Flow) and western blot analysis (WB) using these model systems:
MethodSpeciesCellsTreatmentFixationPerm bufferResult
FlowHumanHeLaS3RNAiBD Cytofix™ III Down-regulation
FlowHumanPBMCNot ApplicableBD Cytofix™ I, II or IIIPositive Staining
WBHumanA431 Cell LysateNot ApplicableNot ApplicableNot Applicable45 kDa
