Orexin-A ELISA Assay Kit

$970.00

The Orexin-A ELISA is used for estimation of Orexin-A in serum, plasma, or tissue extracts in pharmacokinetics, peptide delivery study and other purposes. The Orexin-A ELISA is used for estimation of Orexin-A in serum, plasma, or tissue extracts in pharmacokinetics, peptide delivery study and other purposes.

Orexin-A ELISA Assay Kit

The Orexin-A ELISA Assay KitAssay Kit is For Research Use Only

Size: 96 wells
Standard Range: 40 – 3000 ng/ml
Sensitivity: 40 ng/ml
Incubation Time: 2 hours 30 minutes
Sample Type: Serum, Plasma and Cell Culture Supernatant
Sample Size: 100 µL


Assay Principle

The Orexin-A ELISA is a competitive immunoassay for the determination of Orexin-A. The antiserum is captured by antibodies coated on a 96-well plate. A constant concentration of Biotin Concentrate and varying concentrations of unlabeled standard or sample compete for binding specifically to the antiserum. Captured Bt-tracer is subsequently bound by SA-HRP (streptavidin-conjugated horseradish peroxidase), which produces a soluble colored product after a substrate is added. The enzyme reaction is stopped by dispensing an acidic solution (H2SO4) into the wells after 10 min at room temperature turning the solution from blue to yellow. The optical density (OD) of the solution at 450 nm is inversely proportional to the amount of specific Orexin-A bound. The Orexin-A ELISA is a competitive immunoassay for the determination of Orexin-A. The antiserum is captured by antibodies coated on a 96-well plate. A constant concentration of Biotin Concentrate and varying concentrations of unlabeled standard or sample compete for binding specifically to the antiserum. Captured Bt-tracer is subsequently bound by SA-HRP (streptavidin-conjugated horseradish peroxidase), which produces a soluble colored product after a substrate is added. The enzyme reaction is stopped by dispensing an acidic solution (H2SO4) into the wells after 10 min at room temperature turning the solution from blue to yellow. The optical density (OD) of the solution at 450 nm is inversely proportional to the amount of specific Orexin-A bound.


Specimen Collection and Preparation

Specimens should be clear and non-hemolyzed. Samples should be run at a number of dilutions to ensure accurate quantitation.

Serum: Use a serum separator tube and allow clotting for 30 minutes, then centrifuge for 10 minutes at 1000 x g. Remove serum layer and assay immediately or store serum samples at -20°C. Avoid repeated freeze/thaw cycles.

Dilute the serum 1:50 in Sample Diluent. For example: add 5 ul of serum sample to 245 ul of 1X Sample Diluent. If not assayed immediately, diluted samples should be stored at -20°C or below.


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Additional Information

Assay Background


Orexin-A, also known as hypocretin-1, is a naturally occurring neuropeptide and orexin isoform. The orexinergic nucleus in the lateral hypothalamus is the primary orexin projection system in the brain. Orexin-A is a peptide composed of 33 amino acids including an N-terminal pyroglutamyl residue and two intramolecular disulfide bridges between cysteine residues in 6 and 12 and 7 and 14 positions. Orexin-A, also known as hypocretin-1, is a naturally occurring neuropeptide and orexin isoform. The orexinergic nucleus in the lateral hypothalamus is the primary orexin projection system in the brain. Orexin-A is a peptide composed of 33 amino acids including an N-terminal pyroglutamyl residue and two intramolecular disulfide bridges between cysteine residues in 6 and 12 and 7 and 14 positions.

Documents

Product Manual


Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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