The 5-HETE (5-Hydroxyeicosatetraenoic Acid) ELISA Kit is a highly sensitive and specific assay designed for the accurate detection of 5-HETE levels in a variety of biological samples, including serum, plasma, and cell culture supernatants. This kit offers reliable and reproducible results, making it an essential tool for research applications in the field of lipid metabolism and inflammation.5-HETE is a potent lipid mediator involved in inflammatory responses and immune regulation. It plays a crucial role in various pathological conditions, including cancer, cardiovascular diseases, and inflammatory disorders.
By measuring 5-HETE levels, researchers can gain valuable insights into the mechanisms underlying these diseases and potentially discover new therapeutic targets. Overall, the 5-HETE ELISA Kit provides researchers with a valuable tool for studying the role of 5-HETE in health and disease, ultimately contributing to the development of innovative treatments and interventions.
Product Name:
5-HETE (5-Hydroxyeicosatetraenoic Acid) ELISA Kit
SKU:
UNFI0108
Reactivity:
Universal
Assay Type:
Competitive ELISA, Coated with Antigen
Sensitivity:
0.094 ng/mL
Range:
0.156-10 ng/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)
86-105%
93-102%
92-99%
EDTA Plasma (n = 10)
86-99%
83-90%
85-105%
Heparin Plasma (n = 10)
85-100%
86-99%
80-104%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
85-101
95
EDTA Plasma (n = 10)
88-100
94
Heparin Plasma (n = 10)
88-105
98
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.
Biotin-labeled Antibody (Lyophilized)
1 vial
1 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.
