Secreted Alkaline Phosphatase Reporter Assay Kit

Images

 
ELISA: 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 ...read more

Product Details

Summary
Applications ELISA

Order Details

Secreted Alkaline Phosphatase Reporter Assay Kit Summary

Description
This Kit provides sufficient reagents for 400 quantitative measurements of SEAP protein in 96-well microtiter format. All 96 wells can be used at one time or you may use only the wells as needed by your experimental design. 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.
Immunogen
This Kit provides sufficient reagents for 400 quantitative measurements of SEAP protein in 96-well microtiter format. All 96 wells can be used at one time or you may use only the wells as needed by your experimental design. 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.
Kit Type
Assay Kit

Applications/Dilutions

Application Notes
Sampling Ease: 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.
Publications
Read Publications using
NBP2-25285 in the following applications:

Packaging, Storage & Formulations

Storage
Storage is content dependent.

Kit Components

Components
  1. PNPP Buffer (10X)
  2. PNPP Substrate
  3. Plate Sealer
  4. Dilution Buffer (10X)
  5. 96 well Assay Plates
  6. Secreted Placental Alkaline Phosphatasestandard SEAP (200 ug/ml)

Alternate Names for Secreted Alkaline Phosphatase Reporter Assay Kit

  • SEAP assay

Background

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.

Limitations

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.
Supplier Logo

Publications for Secreted Alkaline Phosphatase Reporter Kit (NBP2-25285)(7)

We have publications tested in 1 confirmed species: Mouse.

We have publications tested in 1 application: ELISA.


Filter By Application
ELISA
(1)
All Applications
Filter By Species
Mouse
(1)
All Species
Showing Publications 1 - 7 of 7.
Publications using NBP2-25285 Applications Species
Fereig RM, Kuroda Y, Terkawi MA et al. Immunization with Toxoplasma gondii peroxiredoxin 1 induces protective immunity against toxoplasmosis in mice. PLoS ONE. Apr 27 2017 [PMID: 28448521] (ELISA, Mouse) ELISA Mouse
Chan M, Hayashi T, Mathewson RD et al. Identification of substituted pyrimido[5,4-b]indoles as selective Toll-like receptor 4 ligands. J Med Chem. 2013 Jun 13 [PMID: 23656327]

Details:
Products cited (Fig 2A): 1. TLR2/NF-kB/SEAPorter HEK 293 cell line (IML-102) 2. TLR3/NF-kB/SEAPorter HEK 293 cell line (IML-103) 3. TLR4/MD2/CD14/NF-kB/SEAPorter HEK 293 cell line (IML-104MD2CD14) 4. TLR5/NF-kB/SEAPorter HEK 293 cell line (IML-105).
Namachivayam K, Blanco CL, Frost BL et al. Preterm human milk contains a large pool of latent TGF-B, which can be activated by exogenous neuraminidase. Am J Physiol Gastrointest Liver Physiol. 2013 Jun 15 [PMID: 23558011]
Potzsch S, Blankenhorn A, Navarrete Santos A et al. The effect of an AGE-rich dietary extract on the activation of NF-kB depends on the cell model used. Food Funct. 2013 Jul [PMID: 23426622]
Ahmed I, Roy B, Chandrakesan P et al. Evidence of functional cross talk between the Notch and NF-kB pathways in nonneoplastic hyperproliferating colonic epithelium. Am J Physiol Gastrointest Liver Physiol. 2013 Feb 15 [PMID: 23203159]

Details:
Products cited (Fig 4B): 1. TLR4/NF-kB/SEAPorter HEK 293 cell line (IML-104) 2. SEAPorter Assay Kit (10055K) Note: Citrobacter rodentium (CR) infected IML-104 TLR4 cells had increased SEAP activity compared to control cells. CR a gram-negative, noninvasiv
Chandrakesan P, Ahmed I, Anwar T et al. Novel changes in NF-{kappa}B activity during progression and regression phases of hyperplasia: role of MEK, ERK, and p38. J Biol Chem. 2010 Oct 22 [PMID: 20710027]

Details:
Products cited (Fig 5A & 5B): TLR4/NF-kB/SEAPorter HEK 293 (IML-104) cell line and the SEAPorter Assay Kit (10055K). Functional Assay: The HEK 293 TLR4 (IML-104) cell line was infected with citrobacter rodentium. SEAP activity was assayed in both the cell
Greene CM, Carroll TP, Smith SG et al. TLR-induced inflammation in cystic fibrosis and non-cystic fibrosis airway epithelial cells. J Immunol. 2005 Feb 1 [PMID: 15661927]

Details:
Products cited (Functional Assays):1.TLR2/NF-kB/SEAP Stably Transfected HEK 293 Cells (IML-102). 2. TLR4/MD-2/NF-kB/SEAP Stably Transfected HEK 293 Cells (IML-104MD2). 3. TLR4/NF-kB/SEAP Stably Transfected HEK 293 Cells (IML-104).4.TLR7/NF-kB/SEAP Stably

Reviews for Secreted Alkaline Phosphatase Reporter Kit (NBP2-25285) (0)

There are no reviews for Secreted Alkaline Phosphatase Reporter Kit (NBP2-25285). By submitting a review you will receive an Amazon e-Gift Card or Novus Product Discount.
  • Review with no image -- $10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen
  • Review with an image -- $25/€18/£15/$25 CAD/¥150 Yuan/¥2500 Yen
  • 1st to review with an image -- $50/€35/£30/$50 CAD/¥300 Yuan/¥5000 Yen

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Secreted Alkaline Phosphatase Reporter Kit (NBP2-25285). (Showing 1 - 1 of 1 FAQs).

  1. 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.
    • The Secreted Alkaline Phosphatase Reporter Kit contains one protein standard. This standard is purified human native protein.

Additional Array Products

Blogs on Secreted Alkaline Phosphatase Reporter

There are no specific blogs for Secreted Alkaline Phosphatase Reporter, but you can read our latest blog posts.

Contact Information

Product PDFs

Review this Product

Be the first to review our Secreted Alkaline Phosphatase Reporter Assay Kit and receive a gift card or discount.