MEK1 antibody

Catalog No. ABIN967763

Product Details anti-MEK1 Antibody

Target: MEK1 (MAP2K1) (Mitogen-Activated Protein Kinase Kinase 1 (MAP2K1))

Reactivity: Human, Mouse, Rat, Dog, Chicken, Frog

Host: Mouse

Clonality: Monoclonal

Conjugate: This MEK1 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), BioImaging (BI)

Cross-Reactivity: Chicken, Dog (Canine), Frog, Mouse (Murine), Rat (Rattus)

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us 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.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human MEK1 Recombinant Protein

Clone: 25-MEK1

Isotype: IgG2a

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl 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 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl 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 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl 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 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN968533, ABIN967389

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-MEK1 antibody (ABIN967763)

Freeman, Brebner, Lynch, Patel, Robertson, Roberts, Vrana: "Changes in rat frontal cortex gene expression following chronic cocaine." in: Brain research. Molecular brain research, Vol. 104, Issue 1, pp. 11-20, (2002) (PubMed).

Gu, Fujibayashi, Yamada, Sekiguchi: "Laminin-10/11 and fibronectin differentially prevent apoptosis induced by serum removal via phosphatidylinositol 3-kinase/Akt- and MEK1/ERK-dependent pathways." in: The Journal of biological chemistry, Vol. 277, Issue 22, pp. 19922-8, (2002) (PubMed).

Aplin, Stewart, Assoian, Juliano: "Integrin-mediated adhesion regulates ERK nuclear translocation and phosphorylation of Elk-1." in: The Journal of cell biology, Vol. 153, Issue 2, pp. 273-82, (2001) (PubMed).

Short, Boyer, Juliano: "Integrins regulate the linkage between upstream and downstream events in G protein-coupled receptor signaling to mitogen-activated protein kinase." in: The Journal of biological chemistry, Vol. 275, Issue 17, pp. 12970-7, (2000) (PubMed).

Robinson, Cheng, Khokhlatchev, Ebert, Ahn, Guan, Stein, Goldsmith, Cobb: "Contributions of the mitogen-activated protein (MAP) kinase backbone and phosphorylation loop to MEK specificity." in: The Journal of biological chemistry, Vol. 271, Issue 47, pp. 29734-9, (1997) (PubMed).

Target Details for MEK1

Target: MEK1 (MAP2K1) (Mitogen-Activated Protein Kinase Kinase 1 (MAP2K1))

Alternative Name: MEK1 (MAP2K1 Products)

Synonyms: Mek1 antibody, CFC3 antibody, MAPKK1 antibody, MEK1 antibody, MKK1 antibody, PRKMK1 antibody, MEKK1 antibody, Prkmk1 antibody, mek-2 antibody, si:ch211-242m18.2 antibody, wu:fj56a12 antibody, wu:fj61b01 antibody, zgc:56557 antibody, ATMEK1 antibody, F20B18.180 antibody, F20B18_180 antibody, MAP kinase/ ERK kinase 1 antibody, MITOGEN ACTIVATED PROTEIN KINASE KINASE antibody, MITOGEN ACTIVATED PROTEIN KINASE KINASE 1 antibody, NMAPKK antibody, MEK antibody, mitogen activated protein kinase kinase 1 antibody, mitogen-activated protein kinase kinase 1 antibody, mitogen-activated protein kinase kinase 1 L homeolog antibody, MAP kinase/ ERK kinase 1 antibody, Map2k1 antibody, MAP2K1 antibody, map2k1.L antibody, map2k1 antibody, MEK1 antibody

Background: 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.

Molecular Weight: 45 kDa

Pathways: MAPK Signaling, RTK Signaling, Interferon-gamma Pathway, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Toll-Like Receptors Cascades, Autophagy, Signaling of Hepatocyte Growth Factor Receptor, BCR Signaling