Q: Could you tell us if Secreted Alkaline Phosphatase Reporter Kit (Cat#NBP2-25285) contains recombinants? If yes, please let us know expression system of the recombinants.

A: The Secreted Alkaline Phosphatase Reporter Kit contains one protein standard. This standard is purified human native protein.

Q: how many 96-well plates can we run with this SEAP assay kit: NBP2-25285

A: This kits comes with 4 plates, 4 plate sealers, and all reagents for 4 x 96-well plates.

Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric)

Novus Biologicals | Catalog # NBP2-25285

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Product Documents

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Product Specifications
Scientific Data
Kit Contents
Preparation & Storage
Background
Product Documents
Specific Notices

Product Specifications

Kit Type

Assay Kit (Colorimetric)

Specificity

Use of duplicate wells for the SEAP standard is recommended to obtain accurate results. This kit can be used with supernatant from transfected cells as described or serum as determined by your experimental protocol.

Description

This Kit provides sufficient reagents for 400 quantitative measurements of SEAP protein in 96-well microtiter format.

Multiple samples can be analyzed in a low-volume, high through-put experiement.

Full analysis complete in under 1 hr.

SEAP is secreted into the media so no cell lysis is required for the detection of its enzymatic activity.

A secreted indicator function permits multiple kinetics experiments using only one culture through sequential sampling of the medium, and also allows the cells to be used for other purposes, such as RNA extraction, Western blot analysis, and other assays.

Heating samples at 65C for 10-30 mins can destroy endogenous alkaline phosphatase but not SEAP.

SEAP is stable in serum, allowing it to be used as a reporter assay in vivo. SEAP catalyzes the hydrolysis of pNitrophenyl phosphate (PNPP) producing a yellow product that can be read in a spectrophotometer or ELISA reader at 405 nm.

Scientific Data Images for Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric)

Enzyme Activity: Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric) [NBP2-25285]

Enzyme Activity: Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric) [NBP2-25285] - Secreted Alkaline Phosphatase Reporter Assay Kit [NBP2-25285] - Standard curve. A serial dilution of SEAP protein was added to wells of a 96-microtiter plate and the standard curve was generated according to the Kit assay protocol.

Kit Contents for Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric)

10X PNPP Buffer (4 mL)
10X Sample Dilution Buffer (10 mL)
96-well Microplate (4 EA)
Microplate Seal (4 EA)
PNPP Substrate (8 x 5 mg)
SEAP Standard at 200 ug/mL (10 uL)

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Storage of components varies. See protocol for specific instructions.

Background: Secreted Alkaline Phosphatase Reporter

Placental alkaline phosphatase (PLAP) is one of the most stable isoenzymes, only existing in the placenta of higher primates. These characteristics make placental alkaline phosphatase suitable to use as a reporter gene for the analysis of promoter activity and gene expression in cell culture and animal serum. The natural form of PLAP is membrane anchored. The recombinant form of placental alkaline phosphatase (secreted alkaline phosphatase, SEAP) is used for reporter gene function. SEAP is designed by inserting a translational terminator after amino acid 489 (Berger, et al., Gene 66 (1): 10 (1988). This mutation converts the membrane-bound PLAP protein into the secreted protein SEAP.

Alternate Names

SEAP assay

Additional Secreted Alkaline Phosphatase Reporter Products

Product Documents for Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric)

Download Product Datasheets

Download SEAP Assay Kit protocol NBP2-25285

Certificate of Analysis

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Product Specific Notices for Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

Citations for Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric)

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