sAPPALPHA ELISA KITS 1-180pg/ml assay ranges (Soluble Amyloid Precursor Protein α, sAPPα; Part No. sAPPalpha-ELISA. Kits for mouse, rat, human, rabbit, canine)

Soluble Amyloid Precursor Protein Alpha High Sensitivity ELISA Kits (sAPPα;  sAPPalpha); Rapid ELISA. 

Separate ELISA kits to sAPPα are available for human, mouse, rat, canine and rabbit sAPPα.  Use the Available Options menu to select the ELISA kit by species.

Fully guaranteed to meet specfications.

• Fully pre-configured sandwich ELISA format providing convenience, high sensitivity and selectivity.
• Strip plate configuration allowing use and storage of separated sections of the ELISA plate.
• Affinity purified polyclonal capture antibody on plate.  Distinct HRP-conjugated. detection antibody.  
  
• Highly sensitive detection of sAPPα in cell culture medium and biological fluids.  Detection ranges are 1-180 pg/ml depending on species.  Detects the soluble alpha-secretase cleavage product of amyloid precursor protein in cell culture medium and biological fluids, designated sAPPα, after centrifugation and sedimentation of cells.  Assays are done with the supernatant fraction to quantitate sAPPα.
  
• This ELISA kit can also detect full length APP (amyloid precursor protein) in cell and tissue lysates after dissolution from the membrane.  To distinguish sAPPα from membrane bound APP, first sediment cells and collect the supernatant to assay sAPPα.  Then use a non-denaturing cell lysis buffer to release and solubilize full length APP from the cell pellet to assay the remaining uncleaved APP. 
  
• Rapid assay with colorimetric detection (measure reaction at 450nm).  Total incubation with ELISA plate and HRP antibody is 1 hr at 37^oC or 2 hr at RT.

Price  (USD)	No. of Kits
$599	1
$1100	2

*  An alternate total sAPP ELISA kit with assay range of 0.39-25ng/ml is also available that uses monoclonal antibody (Cost:  $599).  click here

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sAPPα ELISA Kit Availability And Specifications

Background

sAPPα is the extracellular soluble fragment of amyloid precursor protein cleaved by "alpha-secretase."  Generation of sAPPα by "alpha-secretase" prevents development of fibrillogenic amyloid-beta since the cleavage site is within the amyloid-beta molecule.  The generation of sAPPalpha may therefore be neuroprotective.  sAPPα may function in neuronal excitability, synaptic plasticity, neurite outgrowth, synaptogenesis, and cell survival.  sAPPalpha also appears to modulate potassium channels, N-methyl-D-aspartate receptors, and the transcription factor NF kappa B¹.

Methods Overview:  Sandwich ELISA

The included ELISA 96-well micro-strip plate is provided pre-coated with an affinity purified rabbit polyclonal capture antibody selective to the antigen.  Upon addition of samples to the wells, the antigen in solution binds to the antibody and becomes indirectly linked to the solid support.  After washes of unbound materials, a second affinity purified polyclonal antibody conjugated with horse-radish peroxidase (HRP) is added, followed by a second incubation period.  Unbound antibody is washed, and then color reagents are added which upon conversion by HRP become blue colored.  A stop solution is added to block further reaction between HRP and the colorimetric substrates, converting the solution into a yellow coloration.  An absorbance multiplate reader is used to quantitate the colorimetric reaction at 450 nm.  This assay requires approximately 1.5 hrs to complete at 37^oC, which includes incubation with antibodies, washes and color reaction.

The ELISA assay kit provides a standard peptide solution with known concentration to calibrate absorbance readings to concentration. 

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   Representative Sandwich ELISA Methods                                                                               

Steps of Representative Sandwich ELISA
1.  Equilibrate plate to ambient temperature

2.  Add sample, incubate 30 min - 1hr, wash 

3.  Add HRP-Antibody, incubate 30 min - 1 hr, wash

4.  Add the color reagent, incubate 15 min

5.  Add the stop solution

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Protocol

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Product Citations:

1. Virginie Stygelbout, et al.  2014.  Inositol trisphosphate 3-kinase B is increased in human Alzheimer brain and exacerbates mouse Alzheimer pathology.  Brain, Volume 137, Issue 2  Pp. 537-552.  Link to article 
   
2. Goetzl, E.J., Mustapic, M., Kapogiannis, D., Eitan, E., Lobach, I.V., Goetzl, L., Schwartz, J.B. and Miller, B.L., 2016. Cargo proteins of plasma astrocyte-derived exosomes in Alzheimer’s disease. The FASEB Journal, pp.fj-201600756R.  Link to article
   

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Genereal References on sAPPalpha

1.       Lopez-Perez E, Dumanchin C, Czech C, Campion D, Goud B, et al. 2000. Overexpression of Rab11 or constitutively active Rab11 does not affect sAPPalpha and  Abeta secretions by wild-type and Swedish mutated betaAPP-expressing HEK293 cells. Biochem Biophys Res Commun 275: 910-5

2.       Maillet M, Robert SJ, Cacquevel M, Gastineau M, Vivien D, et al. 2003. Crosstalk between Rap1 and Rac regulates secretion of sAPPalpha. Nat Cell Biol 5: 633-9

3.       Mousavi M, Hellstrom-Lindahl E. 2009. Nicotinic receptor agonists and antagonists increase sAPPalpha secretion and decrease Abeta levels in vitro. Neurochem Int  54: 237-44

4.       Siemes C, Quast T, Klein E, Bieber T, Hooper NM, Herzog V. 2004. Normalized proliferation of normal and psoriatic keratinocytes by suppression of sAPPalpha-release. J Invest Dermatol 123: 556-63

5.       Smirnov A, Trupp A, Henkel AW, Bloch E, Reulbach U, et al. 2009. Differential processing and secretion of Abeta peptides and sAPPalpha in human platelets is regulated by thrombin and prostaglandine 2. Neurobiol Aging 30: 1552-62

6.       Talamagas AA, Efthimiopoulos S, Tsilibary EC, Figueiredo-Pereira ME, Tzinia AK. 2007.

          Abeta(1-40)-induced secretion of matrix metalloproteinase-9 results in sAPPalpha release  by association with cell surface APP. Neurobiol Dis 28: 304-15

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For research only.  Not for diagnostics.

Safety:  Stop solution contains acid.   Avoid ingestion, skin and eye contact.

Storage:  4^oC, 6 months.  Strip plate sections, standard and other solutions can also be stored at -20^oC longer term.

Shipping.  Overnight domestic delivery recommended with styrofoam box/ice pack.  Also available for international delivery; please inquire for shipping cost. 

Kw, sAPPa, sAPP, soluble amyloid precursor, ELISA, alzheimer's, sAPPalpha, sAPPbeta, sAPPb