Immunofluorescent analysis of HeLa cells, untreated (left) or UV-treated (right) using Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) (red). Actin filaments were labeled with Alexa Fluor® 488 phalloidin (green).
|REACTIVITY||H M R Mk Mi Pg Sc|
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 594 fluorescent dye and tested in-house for immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb #4511.
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
NOTE: Cells should be grown, treated, fixed, and stained directly in multi-well plates, chamber slides, or on coverslips.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted Februay 2011
Protocol Id: 6
Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) recognizes endogenous levels of p38 MAPK only when phosphorylated at Thr180 and Tyr182. This antibody does not cross-react with the phosphorylated forms of either p44/42 MAPK or SAPK/JNK.
Human, Mouse, Rat, Monkey, Mink, Pig, S. cerevisiae
Hamster, Chicken, Zebrafish, Bovine, Pig
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr180/Tyr182 of human p38 MAPK protein.
p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAPK, p38α, β, γ (also known as Erk6 or SAPK3), and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAPK is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharide (LPS), UV light, and growth factors (1-5). MKK3, MKK6, and SEK activate p38 MAPK by phosphorylation at Thr180 and Tyr182. Activated p38 MAPK has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6), and MEF2 (5-8). SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole) is a selective inhibitor of p38 MAPK. This compound inhibits the activation of MAPKAPK-2 by p38 MAPK and subsequent phosphorylation of HSP27 (9). SB203580 inhibits p38 MAPK catalytic activity by binding to the ATP-binding pocket, but does not inhibit phosphorylation of p38 MAPK by upstream kinases (10).
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