WB result of Phospho-MEK1 (Ser298) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-MEK1 (Ser298) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HeLa whole cell lysate 20 µg
Lane 2: HeLa starve overnight, then treated with 100 nM Calyculin A for 30 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 43 kDa
Observed MW: 43 kDa
Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho-MEK1 (Ser298) |
| Synonyms | Dual specificity mitogen-activated protein kinase kinase 1; MAP kinase kinase 1; MAPKK 1; MKK1; ERK activator kinase 1; MAPK/ERK kinase 1 (MEK 1); MEK1; PRKMK1; MAP2K1 |
| Immunogen | Synthetic Peptide |
| Location | Cytoplasm, Cytoskeleton, Nucleus |
| Accession | Q02750 |
| Clone Number | S-2487-19 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, ICC |
| Reactivity | Hu, Ms, Rt |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| Dot Blot | 1:1000 | |
| WB | 1:1000-1:2000 | Hu, Ms, Rt |
| ICC | 1:500 | Hu |
Background
Phospho-MEK1 (Ser298) is the activated form of mitogen-activated protein kinase kinase-1 that carries a phosphate group on serine-298; this post-translational modification stabilizes the active conformation of the kinase domain, enabling MEK1 to phosphorylate and activate its only known substrates, ERK1/2, and thereby relay mitogenic, proliferative and differentiation signals from Raf isoforms downstream to diverse cytoplasmic and nuclear targets.
Picture
Picture
Western Blot
WB result of Phospho-MEK1 (Ser298) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody: Phospho-MEK1 (Ser298) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated C6 whole cell lysate 20 µg
Lane 2: C6 starve overnight, then treated with 100 nM Calyculin A for 30 minutes whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 43 kDa
Observed MW: 40 kDa
Dot Blot
Dot blot result of Phospho-MEK1 (Ser298) Recombinant Rabbit mAb
Lane 1: MEK1 (Ser298) phospho peptide
Lane 2: MEK1 (Ser298) unmodified peptide
Primary antibody: Phospho-MEK1 (Ser298) Recombinant Rabbit mAb at 1/1000 dilution
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Immunocytochemistry
ICC analysis of HeLa cells serum-starvation overnight, then treated with Calyculin A (100 nM, 30 min) (top panel) and untreated HeLa cells (below panel). Anti- Phospho-MEK1 (Ser298) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
