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JNK 1/2 (pT183/Y185 + Total) ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the semi-quantitative measurement of JNK 1/2 (pT183/Y185 + Total) Mouse, Human in Tissue Homogenate, Cell Lysate samples.
Colorimetric
Tissue Homogenate, Cell Lysate
Semi-quantitative
Mouse, Human
1h 30m
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
JNK 1/2 (pT183/Y185 + Total) ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the semi-quantitative measurement of JNK 1/2 (pT183/Y185 + Total) Mouse, Human in Tissue Homogenate, Cell Lysate samples.
Colorimetric
Tissue Homogenate, Cell Lysate
Semi-quantitative
Mouse, Human
1h 30m
Microplate
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample (pT183/Y185) | n 6 | mean - | SD - | C.V. 2.7 |
Sample Overall | n 6 | mean - | SD - | C.V. 2.8 |
Sample | n | mean | SD | C.V. |
---|---|---|---|---|
Sample (pT183/Y185) | n 3 | mean - | SD - | C.V. 3.8 |
Sample Overall | n 3 | mean - | SD - | C.V. 3.3 |
Blue Ice
+4°C
+4°C
Abcam's JNK1/2 (pT183/Y185) and JNK1/2 (Total) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of JNK1/2 (pT183/Y185) and Total JNK1/2 protein in Human and mouse cells.
The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.
Estimated sensitivity: Phospho-JNK1/2 (Thr183/Tyr185): 0.5 ng/mL (tested with recombinant protein), Total JNK1/2: 0.1 ng/mL (tested with recombinant protein)
Range: Phospho-JNK1/2 (Thr183/Tyr185): 1-100 ng/mL, Total JNK1/2: 0.2-20 ng/mL
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
The proteins function as critical regulators in cellular processes such as apoptosis inflammation and cell differentiation. JNK1 and JNK2 can form complexes with other proteins to mediate their effects impacting gene expression by phosphorylating transcription factors. These kinases have been extensively studied through techniques like JNK ELISA which measure their activity levels under various conditions. JNK1 and JNK2 are key components in cellular responses to stress and can trigger different outcomes depending on the context and stimuli.
JNK1 and JNK2 also known as Stress-activated Protein Kinases (SAPK) are members of the Mitogen-Activated Protein Kinase (MAPK) family. JNK1 and JNK2 are encoded by the MAPK8 and MAPK9 genes respectively. These kinases are about 46-54 kDa in size and are predominantly expressed in a wide variety of tissues including the brain liver and heart. JNK1 and JNK2 play a central role in transducing signals triggered by stress cytokines and growth factors. They become activated in response to various cellular stresses and are known for their involvement in the regulation of transcription factors such as c-Jun.
The JNK1 and JNK2 proteins integrate into the MAPK signaling cascade which influences diverse cellular activities. They participate in the c-Jun N-terminal kinase pathway an important route affecting the transcription of genes involved in apoptosis. This pathway interacts with proteins such as p53 and ATF2 which further link JNK activity to cellular stress responses. Additionally JNKs contribute to the inflammatory responses by collaborating with NF-kB and other related proteins.
JNK1 and JNK2 have significant roles in the progression of cancer and neurodegenerative disorders like Alzheimer's disease. Aberrant activity or expression of these kinases can lead to enhanced tumorigenesis partly through their interaction with proteins like p53 which regulate cell cycle and apoptosis. In Alzheimer's disease JNKs associate with the pathology by affecting tau phosphorylation linked to neuronal death. Understanding these connections can aid in developing therapeutic strategies targeting JNK pathways.
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Typical cell lysate dilution series.
Example of a typical JNK1/2 (pT183/Y185) and JNK1/2 (Total) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.
Typical recombinant protein standard curve.
Example of a typical JNK1/2 (pT183/Y185) recombinant protein standard curve. The proportion of total protein that is phosphorylated is unknown - data is indicative only. Background-subtracted data values (mean +/- SD) are graphed.
Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of JNK1/2 (pT183/Y185) are normalized and plotted.
Comparison of JNK1/2 expression in different cell lines.
Cell line analysis for Total JNK1/2 from 200 μg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
JNK1/2 (pT183/Y185) phosphorylation in response to anisomycin treatment.
Induction of JNK1/2 (pT183/Y185) phosphorylation in HeLa cells in response to anisomycin treatment. HeLa cells were cultured in 96-well tissue culture plates and treated (30 min) with a dose-range of anisomycin before cell lysis. Data from quadruplicate measurements of JNK1/2 (pT183/Y185) are plotted and compared against Total JNK1/2 protein levels. Comparative JNK1/2 (pT183/Y185) and JNK1/2 (Total) data also shown by Western Blot.
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