The 12-HETE (12-Hydroxyeicosatetraenoic Acid) ELISA Kit is specially designed for the accurate quantification of 12-HETE levels in various sample types, including serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring reliable and reproducible results for a wide range of research applications.12-HETE is a metabolite of arachidonic acid that plays a critical role in inflammation and oxidative stress pathways. Abnormal levels of 12-HETE have been associated with various diseases, including cancer, cardiovascular disorders, and inflammatory conditions.
As such, this ELISA kit serves as a valuable tool for studying the role of 12-HETE in disease development and progression, as well as for developing potential therapeutic interventions. With its user-friendly protocol and high-quality components, the 12-HETE ELISA Kit from Assay Genie is an essential tool for researchers and clinicians interested in investigating the biological functions and clinical implications of 12-HETE in health and disease.
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)
82-98%
86-100%
81-95%
EDTA Plasma (n = 10)
90-98%
87-97%
84-100%
Heparin Plasma (n = 10)
81-97%
84-98%
80-90%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
90-105
93
EDTA Plasma (n = 10)
88-102
96
Heparin Plasma (n = 10)
86-104
97
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.
