Product Pathways - PathScan ELISA
PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit #7946
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|Kit Includes||Volume||Solution Color|
|P-p38 MAPK (T180/Y182) Mouse Ab Coated Microwells||96 tests|
|p38 MAPK Rabbit Detection Antibody||11 ml||Green|
|Anti-rabbit IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
The PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-p38 MAP kinase phosphorylated at Thr180/Tyr182. A phospho-p38 MAP kinase (Thr180/Tyr182) mouse antibody has been coated onto the microwells. After incubation with cell lysates, phospho-p38 MAP kinase (Thr180/Tyr182) protein is captured by the coated antibody. Following extensive washing, a p38 MAP kinase rabbit detection antibody is added to detect the captured phospho-p38 MAP kinase (Thr180/Tyr182). HRP-linked, anti-rabbit antibody is then used to recognize the bound detection antibody. The HRP substrate TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-p38 MAP kinase (Thr180/Tyr182).Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit detects endogenous levels of p38 MAP kinase phosphorylated at Thr180/Tyr182 in human and mouse cells. As shown in Figure 1, a significant induction of p38 MAP kinase phosphorylation at Thr180/Tyr182 can be detected in HeLa cells using the PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit following treatment with anisomycin or UV irradiation. The level of total p38 MAP kinase remains unchanged as shown by western analysis (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1. Treatment of HeLa cells with anisomycin or UV irradiation stimulates phosphorylation of p38 MAP kinase at Thr180/Tyr182 as detected by the PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit #7946, but does not affect the level of total p38 MAP kinase. HeLa cells were treated with 5 μg/ml anisomycin for 30 minutes at 37ºC or 45 mJ/cm2 UV irradiation followed by a 30 minute recovery period at 37ºC. The absorbance readings at 450 nm are shown in the top figure, while the western blots using p38 MAPK Antibody #9212 (left) or Phospho-p38 MAPK (Thr180/Tyr182) Antibody #9211 (right) are shown in the bottom figure.
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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- Kumar, S. et al. (1999) Biochem Biophys Res Commun 263, 825-31.
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- 4511 Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP® Rabbit mAb
- 9215 Phospho-p38 MAPK (Thr180/Tyr182) (3D7) Rabbit mAb
- 4631 Phospho-p38 MAPK (Thr180/Tyr182) (12F8) Rabbit mAb
- 9216 Phospho-p38 MAPK (Thr180/Tyr182) (28B10) Mouse mAb
- 8690 p38 MAPK (D13E1) XP® Rabbit mAb
- 9212 p38 MAPK Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
For Research Use Only. Not For Use In Diagnostic Procedures.