The Platelet-Activating Factor (PAF) ELISA Kit is a cutting-edge tool designed for the precise detection of PAF levels in biological samples such as serum, plasma, and cell culture supernatants. With superior sensitivity and specificity, this kit delivers accurate and consistent results, making it an invaluable asset for a variety of research endeavors. PAF is a bioactive lipid molecule known for its pivotal role in inflammation, allergic responses, and immune regulation.
By measuring PAF levels, researchers can gain valuable insights into the mechanisms underlying various pathological conditions, including asthma, cardiovascular diseases, and inflammatory disorders. This ELISA kit thus presents a powerful resource for studying PAF-related pathologies and facilitating the development of targeted therapeutic interventions.
Product Name:
PAF (Platelet Activating Factor) ELISA Kit
SKU:
UNFI0061
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)
88-100%
85-94%
82-98%
EDTA Plasma (n = 10)
80-98%
85-97%
82-92%
Heparin Plasma (n = 10)
80-98%
80-100%
81-96%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
87-105
95
EDTA Plasma (n = 10)
85-105
98
Heparin Plasma (n = 10)
86-105
95
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.
HRP-Streptavidin Conjugate (SABC, 100X)
60 ul
120 ul
2-8°C (Avoid direct light)
TMB Substrate
5 ml
10 ml
2-8°C (Avoid direct light)
Purified Water
200 ul
200 ul
2-8°C
Sample Dilution Buffer
10 ml
20 ml
2-8°C
Antibody Dilution Buffer
5 ml
10 ml
2-8°C
SABC Dilution Buffer
5 ml
10 ml
2-8°C
Stop Solution
5 ml
5 ml
2-8°C
Wash Buffer(25X)
15 ml
30 ml
2-8°C
Plate Sealer
3 pieces
5 pieces
-
Technical Manual
1 copy
1 copy
-
Carolyn J. English et al.
Novel inflammatory markers PAF and Lp-PLA2 in cardiovascular disease prevention: relationships with dietary patterns and traditional risk factors
