Key features and details
- One-wash 90 minute protocol
- Sample type: Cell Lysate, Tissue Homogenate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Mouse, Human
Product nameJNK 1/2 (pT183/Y185 + Total) ELISA Kit
Intra-assay Sample n Mean SD CV% (pT183/Y185) 6 2.7% (Total) 6 2.8% Inter-assay Sample n Mean SD CV% (pT183/Y185) 3 3.8% (Total) 3 3.3%
Sample typeCell Lysate, Tissue Homogenate
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat
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.
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Wash Buffer PT 1 x 15ml 50X Cell Extraction Enhancer Solution 1 x 1ml 5X Cell Extraction Buffer PTR 1 x 12ml JNK1/2 (pT183/Y185) Capture Antibody 1 x 1.5ml JNK1/2 (pT183/Y185) Detector Antibody 1 x 1.5ml JNK1/2 (Total) Capture Antibody 1 x 1.5ml JNK1/2 (Total) Detector Antibody 1 x 1.5ml Lyophilized JNK1/2 Control Lysate 1 vial Plate Seal 1 unit SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Substrate 1 x 12ml
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
Example of a typical JNK1/2 (pT183/Y185) and JNK1/2 (Total) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.
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.
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).
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.
ab176662 has been referenced in 4 publications.
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- Caldwell CC et al. Treadmill exercise rescues mitochondrial function and motor behavior in the CAG140 knock-in mouse model of Huntington's disease. Chem Biol Interact 315:108907 (2019). PubMed: 31778667
- Tian L et al. Calotropin regulates the apoptosis of non-small cell cancer by regulating the cytotoxic T-lymphocyte associated antigen 4-mediated TGF-ß/ERK signaling pathway. Mol Med Rep 17:7683-7691 (2018). PubMed: 29620207
- Chen X et al. Evodiamine alleviates severe pneumonia induced by methicillin-susceptible Staphylococcus aureus following cytomegalovirus reactivation through suppressing NF-?B and MAPKs. Int J Mol Med 42:3247-3255 (2018). PubMed: 30320369