The Human 12-HETE ELISA Kit is specifically developed for the precise measurement of 12-Hydroxyeicosatetraenoic acid (12-HETE) levels in human samples such as serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, guaranteeing accurate and consistent results, making it an excellent choice for various research purposes. 12-HETE is a lipid mediator that is involved in numerous biological processes, including inflammation, oxidative stress, and cell proliferation. Elevated levels of 12-HETE have been linked to various diseases such as cancer, cardiovascular disorders, and inflammatory conditions, highlighting its significance as a potential biomarker for disease progression and treatment monitoring.
With the Human 12-HETE ELISA Kit, researchers can efficiently study the role of 12-HETE in disease pathogenesis and potentially identify novel therapeutic targets for the development of targeted therapies. Trust in the reliability and precision of this kit for accurate measurement of 12-HETE levels in human samples.
Product Name:
Human 12-HETE ELISA Kit
SKU:
HUFI01660
Reactivity:
Human
Assay Type:
Competitive ELISA, Coated with Antigen
Sensitivity:
0.188 ng/mL
Range:
0.313-20 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)
85-96%
91-100%
86-104%
EDTA Plasma (n = 10)
88-100%
87-95%
82-100%
Heparin Plasma (n = 10)
94-100%
86-95%
80-100%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
85-102
96
EDTA Plasma (n = 10)
93-105
100
Heparin Plasma (n = 10)
87-102
93
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.
