Skip to main content Skip to navigation
Purified Mouse Anti-MEK1
Purified Mouse Anti-MEK1
Western blot analysis of MEK1 on a A431 lysate (Cat. No 611447). Lane 1: 1:1000, lane 2: 1:2000, lane 3: 1:4000 dilution of the anti-MEK1 antibody.
Purified Mouse Anti-MEK1
Immunofluorescent staining of HeLa cells (ATCC CCL-2).  Cells were seeded in a 96 well imaging plate (Cat. No. 353219) at ~10,000 cells per well. After overnight incubation, cells were stained using the alcohol perm protocol and the anti-MEK1 antibody.  The second step reagent was FITC goat anti mouse Ig (Cat. No. 554001). The image was taken on a BD Pathway™ 855 Bioimager using a 20x objective. This antibody also stained A549 (ATCC CCL-185) and U2OS (ATCC HTB-96) cells and can be used with either perm protocol (see Recommended Assay Procedure).
Western blot analysis of MEK1 on a A431 lysate (Cat. No 611447). Lane 1: 1:1000, lane 2: 1:2000, lane 3: 1:4000 dilution of the anti-MEK1 antibody.
Immunofluorescent staining of HeLa cells (ATCC CCL-2).  Cells were seeded in a 96 well imaging plate (Cat. No. 353219) at ~10,000 cells per well. After overnight incubation, cells were stained using the alcohol perm protocol and the anti-MEK1 antibody.  The second step reagent was FITC goat anti mouse Ig (Cat. No. 554001). The image was taken on a BD Pathway™ 855 Bioimager using a 20x objective. This antibody also stained A549 (ATCC CCL-185) and U2OS (ATCC HTB-96) cells and can be used with either perm protocol (see Recommended Assay Procedure).
Product Details
Down Arrow Up Arrow


BD Transduction Laboratories™
Human (QC Testing), Mouse,Rat,Dog,Chicken,Frog (Tested in Development)
Mouse IgG2a
Human MEK1 Recombinant Protein
Western blot (Routinely Tested), Bioimaging, Immunohistochemistry, Immunoprecipitation (Tested During Development)
45 kDa
250 µg/ml
AB_397528
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at -20°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Bioimaging

1. Seed the cells in appropriate culture medium at ~10,000 cells per well in a BD Falcon™ 96-well Imaging Plate (Cat. No. 353219) and culture overnight.

2. Remove the culture medium from the wells, and fix the cells by adding 100 μl of BD Cytofix™ Fixation Buffer (Cat. No. 554655) to each well.  Incubate for 10 minutes at room temperature (RT).

3. Remove the fixative from the wells, and permeabilize the cells using either BD Perm Buffer III, 90% methanol, or Triton™ X-100:

a. Add 100 μl of -20°C 90% methanol or Perm Buffer III (Cat. No. 558050) to each well and incubate for 5 minutes at RT.

     OR

b. Add 100 μl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.

4. Remove the permeabilization buffer, and wash the wells twice with 100 μl of 1× PBS.

5. Remove the PBS, and block the cells by adding 100 μl of BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) to each well. Incubate for 30 minutes at RT.

6. Remove the blocking buffer and add 50 μl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.

7. Remove the primary antibody, and wash the wells three times with 100 μl of 1× PBS.

8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 μl to each well, and incubate in the dark for 1 hour at RT.

9. Remove the second step reagent, and wash the wells three times with 100 μl of 1× PBS.

10. Remove the PBS, and counter-stain the nuclei by adding 200 μl per well of 2 μg/ml Hoechst 33342 (e.g., Sigma-Aldrich Cat. No. B2261) in 1× PBS to each well at least 15 minutes before imaging.

11. View and analyze the cells on an appropriate imaging instrument.

Bioimaging:  For more detailed information please refer to http://www.bdbiosciences.com/support/resources/protocols/ceritifed_reagents.jsp

Western blot:  For more detailed information please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
  4. 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.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. Triton is a trademark of the Dow Chemical Company.
610122 Rev. 2
Antibody Details
Down Arrow Up Arrow
25/MEK1

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.

610122 Rev. 2
Format Details
Down Arrow Up Arrow
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
610122 Rev.2
Citations & References
Down Arrow Up Arrow
View product citations for antibody "610122" on CiteAb

Development References (5)

  1. Aplin AE, Stewart SA, Assoian RK, Juliano RL. Integrin-mediated adhesion regulates ERK nuclear translocation and phosphorylation of Elk-1. J Cell Biol. 2001; 153(2):273-282. (Clone-specific: Fluorescence microscopy, Immunofluorescence, Western blot). View Reference
  2. Freeman WM, Brebner K, Lynch WJ, et al. Changes in rat frontal cortex gene expression following chronic cocaine. Brain Res Mol Brain Res. 2002; 104(1):11-20. (Clone-specific: Western blot). View Reference
  3. Gu J, Fujibayashi A, Yamada KM, Sekiguchi K. Laminin-10/11 and fibronectin differentially prevent apoptosis induced by serum removal via phosphatidylinositol 3-kinase/Akt- and MEK1/ERK-dependent pathways. J Biol Chem. 2002; 277(22):19922-19928. (Clone-specific: Western blot). View Reference
  4. Robinson MJ, Cheng M, Khokhlatchev A, et al. Contributions of the mitogen-activated protein (MAP) kinase backbone and phosphorylation loop to MEK specificity. J Biol Chem. 1996; 271(47):29734-29739. (Biology). View Reference
  5. Short SM, Boyer JL, Juliano RL. Integrins regulate the linkage between upstream and downstream events in G protein-coupled receptor signaling to mitogen-activated protein kinase. J Biol Chem. 2000; 275(17):12970-12977. (Clone-specific: Immunoprecipitation, In vitro kinase assay, Western blot). View Reference
View All (5) View Less
610122 Rev. 2

Please refer to Support Documents for Quality Certificates


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.