CREB1 pS133 + CREB1 ELISA Kit (ab176659)
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
Overview
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Product name
CREB1 pS133 + CREB1 ELISA Kit -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% CREB1(pS133) 6 4.7% CREB1(Total) 6 4.1% Inter-assay Sample n Mean SD CV% CREB1(pS133) 3 3.9% CREB1(Total) 3 4.5% -
Sample type
Cell Lysate, Tissue Homogenate -
Assay type
Semi-quantitative -
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 CREB1 (pS133) and CREB1 (Total) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of CREB1 (pS133) and Total CREB1 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
Estimated sensitivity: Phospho-CREB (Ser133): 0.5 µg/mL (tested with MCF-7), Total CREB: 1.0 µg/mL (tested with MCF-7)
Range: Phospho-CREB (Ser133): 1-100 µg/mL, Total CREB: 2-200 µg/mLAbcam 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 CREB1 (pS133) Capture Antibody 1 x 1.5ml CREB1 (pS133) Detector Antibody 1 x 1.5ml CREB1 (Total) Capture Antibody 1 x 1.5ml CREB1 (Total) Detector Antibody 1 x 1.5ml Lyophilized CREB1 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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Cellular localization
CREB: Nucleus. CREB1 (Total): Nucleus. -
Database links
- Entrez Gene: 1385 Human
- Entrez Gene: 12912 Mouse
- Entrez Gene: 12912 Mouse
- Entrez Gene: 81646 Rat
- Entrez Gene: 81646 Rat
- Omim: 123810 Human
- Omim: 123810 Human
- SwissProt: P16220 Human
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Images
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Other - CREB1 pS133 + CREB1 ELISA Kit (ab176659)
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 CREB1 (pS133) and CREB1 (Total) cell lysate dilution series. 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 CREB1 (pS133) are normalized and plotted.
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Comparison of CREB1 expression in different cell linesCell line analysis for Total CREB1 from 100 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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CREB1 (pS133) phosphorylation in response to AG1486 treatmentInhibition of CREB1 (pS133) phosphorylation in MCF-7 cells in response to AG1486 treatment. HeLa cells were cultured in 96-well tissue culture plates and treated (60 min) with a dose-range of AG1486. Cells were then stimulated (10 min) with 100 ng/mL EGF and lysed. Data from quadruplicate measurements of CREB1 (pS133) are plotted and compared against total CREB1 protein levels. Comparative CREB1 (pS133) and CREB1 (Total) data also shown by Western Blot.
