p38α (Thr180/Tyr182) Dual Detect CELISA Assay Kit (Fluorogenic Detection)
Description:
p38α (Thr180/Tyr182) Dual Detect CELISA Assay Kit (Fluorogenic Detection)
Trade Name:
Dual Detect CELISA
Product Overview:
Dual Detect CELISA is a straightforward and efficient cell-based ELISA assay that eliminates the need to prepare cell lysates. It allows for the measurement of the relative expression levels of both the total and the phosphorylated forms of the target protein in whole cells in a convenient 96 well microplate format using dual fluorescent detection. This allows for the normalization of total protein between wells, thus allowing a more accurate measurement of phosphorylation levels among cell and/or growth condition differentials.
Cells are cultured directly in the microplate and stimulated as desired. They are then fixed and simultaneously incubated with two primary antibodies. One is a phosphospecific antibody directed against the phosphorylated epitope of the protein that correlates with it activation and the other is a total antibody that recognizes the target-protein, regardless of phosphorylation status. the levels of both the total target protein and the phosphorylated protein are measured simultaneously in a single well using a double immunoenzymatic labeling procedure, either horseradish-peroxidase (HRP) or alkaline phosphatase (AP), and two spectrally distinct fluorogenic substrates for HRP and AP. The fluorescence of the phosphorylated protein is normalized to the total protein in each well thus reducing well-to-well variations. The p38 (Thr180/Tyr182) kit contains the components required to run the Dual Detect CELISA using fluorogenic substrates to measure both total p38 and phosphorylated p38 (Thr180/Tyr182) in the context of whole cells.
Cells are cultured directly in the microplate and stimulated as desired. They are then fixed and simultaneously incubated with two primary antibodies. One is a phosphospecific antibody directed against the phosphorylated epitope of the protein that correlates with it activation and the other is a total antibody that recognizes the target-protein, regardless of phosphorylation status. the levels of both the total target protein and the phosphorylated protein are measured simultaneously in a single well using a double immunoenzymatic labeling procedure, either horseradish-peroxidase (HRP) or alkaline phosphatase (AP), and two spectrally distinct fluorogenic substrates for HRP and AP. The fluorescence of the phosphorylated protein is normalized to the total protein in each well thus reducing well-to-well variations. The p38 (Thr180/Tyr182) kit contains the components required to run the Dual Detect CELISA using fluorogenic substrates to measure both total p38 and phosphorylated p38 (Thr180/Tyr182) in the context of whole cells.
Background Information:
The p38 family of kinases, also known as the SAPK2/3 Kinases, are a subfamily of JNK/SAPK family that have traditionally been associated with stress, immune response, and regulation of cell proliferation. The p38 MAPK family of Ser/Thr kinases, referred to as the stress-activated MAPK, is made up of 4 isoforms, p38(MAPK14, SAPK2a), p38(MAPK11, SAPK2b), p38(MAPK13, SAPK4), and p38(MAPK12, SAPK3). They are regulated by stress-inducing signals such as osmotic stress, UV radiation, pro-inflammatory cytokines such as IL-1 and TNF, and certain toxins such as anisomycin. Upon stimulation, they are dually phosphorylated on their common TGY motif resulting in their activation, which is Thr180/Tyr182 in human p38. Once activated, p38 is able to phosphorylate a wide array of targets including numerous kinases such as MAPKAP-K2, MAPKAP-K3, PRAK, MSK, and MNK and many transcription factors including STAT1, p53, NFATp, ATF-2, and MEF2. p38 has roles in regulating the production of pro-inflammatory cytokines and has been implicated in many pathological conditions that include inflammation, rheumatoid arthritis, diabetes, cancer, and neurodegenerative diseases.
Species Reactivity:
- Human
- Mouse
- Rat
Alternate Names:
- SAPK2a
- MAPK14
Modifications:
Phosphorylation
Components:
- 500X anti-p38 (Total) Antibody
- 500X anti-phospho p38 (Thr180/Tyr182) Antibody
- 500X Goat anti-Mouse HRP Detection Reagent
- 500X Goat anti-Rabbit AP Detection Reagent
- 200X HRP Substrate
- 200X AP Substrate
- Black Clear Bottom 96-well Cell Culture Plate
- 20% Tween® 20 (v/v)
- 20X TBS
- 10% BSA in TBS (Blocking Buffer)
- Substrate Diluent
Kit Type:
- Cell-based ELISA
- CELISA
Gene Symbol:
- MAPK14
- CSBP
- CSBP1
- CSPB1
- MXI2
- SAPK2A
Analytes Available:
p38
Quality Assurance:
Routinely QC tested
Detection Methods:
Fluorescent
Kit or Assay Type:
- Cell-based ELISA
- CELISA
- Cell Based Assays
- ELISA Assays
- Kinase & Phosphatase Assays
- Signal Transduction Assays
Assay Type:
- Cell Based Assays
- ELISA Assays
- Kinase & Phosphatase Assays
- Signal Transduction Assays
Storage Conditions:
-20C