Phospho-ERK1/ERK2/JNK1/P38 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-ERK1/ERK2/JNK1/P38 in Human in Cell Lysate samples.
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Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Phospho-ERK1/ERK2/JNK1/P38 ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-ERK1/ERK2/JNK1/P38 in Human in Cell Lysate samples.
Phospho-ERK1/ERK2/JNK1/P38 ELISA Kit (ab279855) is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated ERK1/2, JNK and P38 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.
This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-ERK1/2, total ERK1/2, phospho-JNK, total JNK, phospho- P38 and total P38. For each target, a capture antibody has been coated onto microwells. Samples are pipetted into the wells and target protein present in a sample is bound to the wells by the immobilized antibody. The wells are washed and a detection antibody is used to detect the captured target protein. After washing away unbound antibody, an HRP-conjugated secondary antibody is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of target protein bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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The proteins p38 JNK1 ERK1 and ERK2 are critical members of the mitogen-activated protein kinase (MAPK) family. These kinases play key roles in cellular signaling. p38 also known as MAPK14 typically has a mass of approximately 38 kDa. It is expressed in various tissues including the lung liver and muscle. JNK1 or MAPK8 with a mass around 46 kDa is expressed in many tissues with high levels in the brain and heart. ERK1 (MAPK3) and ERK2 (MAPK1) have masses of about 44 kDa and 42 kDa respectively and show wide expression particularly in cells with high mitotic activity such as epithelial cells. Despite their different sizes they share similarities in their sequences and activities.
These MAPKs contribute to cell cycle regulation apoptosis differentiation and stress responses. p38 functions in stress-induced responses and inflammation while JNK1 plays a role in apoptosis and neurodegeneration. ERK1 and ERK2 are part of the ERK pathway influencing cell proliferation and differentiation. These proteins often operate as part of larger signaling complexes such as the p38 MAPK cascade or the JNK signal transduction network which allow cells to respond accurately to various extracellular stimuli. Their activity often involves phosphorylation leading to subsequent activation or inhibition of downstream effector proteins.
These MAPKs serve prominent roles in the regulation of the MAPK signaling pathways specifically the p38 MAPK and ERK pathways. The p38 MAPK pathway facilitates responses to stress stimuli like cytokines and UV radiation while the ERK pathway is critical for growth factor signaling. Within these pathways these proteins can interact with upstream activators like MEK1 and MEK2 which phosphorylate and activate them. They can also phosphorylate downstream targets such as transcription factors which can modify gene expression patterns in response to environmental changes.
These MAPKs are linked to conditions such as rheumatoid arthritis and cancer. p38 is associated with inflammatory diseases like rheumatoid arthritis due to its role in producing pro-inflammatory cytokines. Researchers find that targeting p38 can potentially alleviate symptoms in such disorders. JNK1 and its role in apoptosis connect it to neurodegenerative diseases such as Alzheimer's disease. ERK1 and ERK2 through their involvement in the regulation of cell proliferation relate strongly to cancer progression alongside proteins like the tumor suppressor p53. Understanding these connections is important for developing therapeutic strategies targeting these pathways.
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Positive Control.
A431 cells were treated with recombinant human EGF at 37°C for 20 mins.
Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer.
Serial dilutions of lysates were analyzed using phospho-ERK1/2 strips in this ELISA.
HeLa cells treated with/without Anisomycin.
HeLa cells were treated or untreated with Anisomycin for 10 minutes at 37°C.
HeLa cells treated with/without Anisomycin.
HeLa cells were treated or untreated with Anisomycin for 10 minutes at 37°C.
HepG2 cells untreated/treated with IL-1 beta.
HepG2 cells were treated or untreated with 25ng/ml IL-1 beta for 30 minutes.
A431 cells treated with/without EGF. A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 20 minutes.
A431 cells treated with/without EGF. A431 cells were treated or untreated with 100 ng/ml recombinant human EGF for 20 minutes.
HepG2 cells untreated/treated with IL-1 beta.
HepG2 cells were treated or untreated with 25ng/ml IL-1 beta for 30 minutes.
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