The Sheep MT (Melatonin) ELISA Kit is specifically designed for the quantitative detection of melatonin levels in sheep serum, plasma, and tissue homogenates. This kit offers high sensitivity and specificity, ensuring accurate and consistent results for researchers in the field of chronobiology and neuroendocrinology. Melatonin is a hormone produced by the pineal gland that regulates the sleep-wake cycle and circadian rhythms in animals.
It also has antioxidant and anti-inflammatory properties, making it a key player in various physiological processes. Studies have shown that melatonin levels can be affected by factors such as age, light exposure, and health conditions, making this kit an invaluable tool for studying melatonin dynamics in sheep and its potential implications in sleep disorders, seasonal changes, and overall health.
Product Name:
Sheep MT (Melatonin) ELISA Kit
SKU:
SHFI00052
Reactivity:
Sheep
Assay Type:
Competitive ELISA, Coated with Antigen
Sensitivity:
4.688 pg/mL
Range:
7.813-500 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)
88-102%
85-99%
80-95%
EDTA Plasma (n = 10)
88-98%
87-99%
81-94%
Heparin Plasma (n = 10)
87-101%
83-101%
85-99%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
85-105
94
EDTA Plasma (n = 10)
85-104
95
Heparin Plasma (n = 10)
87-102
96
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.
