The Rat Orexin-A ELISA Kit is a highly sensitive and specific assay designed for the precise measurement of orexin-A levels in rat serum, plasma, and cell culture supernatants. This kit offers reliable and reproducible results, making it suitable for a variety of research applications. Orexin-A, also known as hypocretin-1, is a neuropeptide that plays a crucial role in regulating wakefulness and sleep. Dysfunction of the orexin system has been linked to various sleep disorders, including narcolepsy and insomnia.
By accurately measuring orexin-A levels, researchers can better understand the underlying mechanisms of these disorders and develop potential therapeutic interventions. Overall, the Rat Orexin-A ELISA Kit is a valuable tool for studying the orexin system and its implications in sleep-wake regulation, making it an essential component of any research focused on sleep disorders and related conditions in rats.
Product Name:
Rat Orexin A ELISA Kit
SKU:
RTFI01016
Reactivity:
Rat
Assay Type:
Competitive ELISA, Coated with Antigen
Sensitivity:
37.5 pg/mL
Range:
62.5-4000 pg/mL
Sample Type:
Serum, Plasma, Cell Culture Supernatant, Cell or Tissue Lysate, Other Liquid Samples
Storage:
2-8°C(Sealed), Don't cryopreserve.
Linearity:
Sample
1:2
1:4
1:8
Serum (n = 10)
90-105%
85-100%
88-101%
EDTA Plasma (n = 10)
97-101%
83-100%
84-98%
Heparin Plasma (n = 10)
90-100%
85-98%
87-93%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
86-99
93
EDTA Plasma (n = 10)
87-100
93
Heparin Plasma (n = 10)
91-105
99
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Step
Procedure
1
Reagent & Plate Preparation: Equilibrate TMB substrate for 30 minutes at room temperature. Prepare standards, samples (minimum 1:2 dilution), blanks, assign wells, and pre-wash the plate twice.
2
Sample & Biotin-Antibody Binding: Add 50 µL standard or sample followed by 50 µL biotin-labeled antibody to each well. Mix gently and incubate at 37°C for 45 minutes.
3
Washing: Wash the plate 5 times with wash buffer, allowing 1 minute soak time per wash.
4
Color Development: Add TMB substrate and incubate in the dark at 37°C for 10-20 minutes until color develops.
5
Stop Reaction: Add stop solution to terminate the reaction. The color changes from blue to yellow immediately.
6
Reading: Measure absorbance at 450 nm using a microplate reader.
Sample Type
Protocol
Serum
Allow blood to clot, centrifuge at 1000 × g for 20 minutes, collect supernatant and store appropriately.
Plasma
Collect using EDTA anticoagulant, centrifuge at 1000 × g for 15 minutes at 2–8°C and collect plasma.
Cell Culture Supernatant
Centrifuge at 1000 × g for 20 minutes at 4°C and collect clarified supernatant.
Cell Lysate
Lyse cells using recommended lysis buffer with protease inhibitors, centrifuge at 10,000 rpm for 10 minutes, and collect protein supernatant.
Tissue Homogenate
Homogenize tissue in PBS with protease inhibitors, centrifuge at 5000 × g for 5 minutes, and collect supernatant.
Other Sample Types
Centrifuge samples at 1000 × g for 15 minutes at 2–8°C and collect supernatant. For additional guidance, please contact techsupport@assaygenie.com.
Component
Quantity
Storage
48T
96T
ELISA Microplate (Dismountable)
8×6
8×12
Place the test strips into a sealed foil bag with the desiccant. Store for 1 month at 2-8°C; Store for 12 months at -20°C.
Lyophilized Standard
1 vial
2 vial
Place the standards into a sealed foil bag with the desiccant. Store for 1 month at 2-8°C; Store for 12 months at -20°C.
system_update_altDatasheetRat Orexin (OX) ELISA KitThe Rat Orexin (OX) ELISA Kit is a top-quality assay kit developed for the precise measurement of orexin levels in rat sampl
