JNK 1/2 ELISA Kit (ab176646)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 0.1 ng/ml
- Range: 0.2 ng/ml - 20 ng/ml
- Sample type: Cell Lysate, Tissue Homogenate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Mouse, Human
Overview
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Product name
JNK 1/2 ELISA Kit -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% HEK extracts 6 2.8% Inter-assay Sample n Mean SD CV% HEK extracts 3 3.3% -
Sample type
Cell Lysate, Tissue Homogenate -
Assay type
Semi-quantitative -
Sensitivity
0.1 ng/ml -
Range
0.2 ng/ml - 20 ng/ml -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat
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Product overview
Abcam’s JNK1/2 in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of 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.
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Notes
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. -
Platform
Microplate
Properties
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Storage instructions
Store 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 (Total) Capture Antibody 1 x 3ml JNK1/2 (Total) Detector Antibody 1 x 3ml 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 -
Research areas
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Relevance
Responds to activation by environmental stress and pro-inflammatory cytokines by phosphorylating a number of transcription factors, primarily components of AP-1 such as JUN, JDP2 and ATF2 and thus regulates AP-1 transcriptional activity. In T-cells, JNK1 and JNK2 are required for polarized differentiation of T-helper cells into Th1 cells (By similarity). Phosphorylates heat shock factor protein 4 (HSF4). JNK1 isoforms display different binding patterns: beta-1 preferentially binds to c-Jun, whereas alpha-1, alpha-2, and beta-2 have a similar low level of binding to both c-Jun or ATF2. However, there is no correlation between binding and phosphorylation, which is achieved at about the same efficiency by all isoforms. -
Database links
- Entrez Gene: 5599 Human
- Entrez Gene: 26419 Mouse
- Entrez Gene: 116554 Rat
- SwissProt: P45983 Human
- SwissProt: Q91Y86 Mouse
- SwissProt: P49185 Rat
- Unigene: 138211 Human
- Unigene: 21495 Mouse
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Images
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Other - JNK 1/2 ELISA Kit (ab176646)
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.
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Typical cell lysate dilution seriesExample of a typical JNK1/2 cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.
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Typical recombinant protein standard curveExample of a typical JNK1/2 recombinant protein standard curve. Background-subtracted data values (mean +/- SD) are graphed.
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Linearity of dilutionLinearity 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 are normalized and plotted.
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Comparison of JNK 1/2 expression in different cell linesCell 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).
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JNK 1/2 phosphorylation in response to anisomycin treatmentInduction 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.
