Human 5-HETE/5-Hydroxyeicosatetraenoic Acid ELISA Kit
The Human 5-HETE (5-hydroxyeicosatetraenoic acid) ELISA Kit is specifically designed for the precise measurement of 5-HETE levels in human samples such as serum, plasma, and cell culture supernatants. This kit offers unparalleled sensitivity and specificity, guaranteeing accurate and consistent results, making it an invaluable tool for various research applications.5-HETE is a key metabolite of arachidonic acid and plays a critical role in inflammation, oxidative stress, and various disease processes. Its dysregulation has been implicated in numerous pathological conditions, including cancer, cardiovascular diseases, and inflammatory disorders, highlighting its significance as a biomarker for understanding and potentially treating these diseases.
With the Human 5-HETE ELISA Kit, researchers can precisely quantify 5-HETE levels in biological samples, furthering our understanding of its role in health and disease. This kit provides a reliable and efficient method for studying 5-HETE metabolism and its implications in various pathological conditions, paving the way for future therapeutic interventions.
Product Name:
Human 5-HETE/5-Hydroxyeicosatetraenoic Acid ELISA Kit
SKU:
HUFI01774
Reactivity:
Human
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)
88-101%
94-105%
87-105%
EDTA Plasma (n = 10)
82-92%
83-98%
92-99%
Heparin Plasma (n = 10)
83-98%
80-97%
80-99%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
88-100
97
EDTA Plasma (n = 10)
85-99
92
Heparin Plasma (n = 10)
89-105
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.
