The Canine TNF-beta (Tumor Necrosis Factor-beta) ELISA Kit is specifically designed for the accurate measurement of TNF-beta levels in canine serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring reliable and reproducible results for a variety of research applications. TNF-beta is a key cytokine involved in inflammatory processes, immune response modulation, and cell apoptosis. Dysregulation of TNF-beta has been implicated in various diseases, including autoimmune disorders, infectious diseases, and cancer.
Therefore, this ELISA kit serves as a valuable tool for studying TNF-beta biology and its role in disease pathogenesis. Overall, the Canine TNF-beta ELISA Kit from Assay Genie provides researchers with a precise and efficient method for quantifying TNF-beta levels in canine samples, facilitating in-depth investigations into the mechanisms underlying inflammatory and immune-related conditions in dogs.
Serum, Plasma, Cell Culture Supernatant, Cell or Tissue Lysate, Other Liquid Samples
Storage:
2-8°C for 12 months.
Linearity:
Sample
1:2
1:4
1:8
Serum (n = 5)
82-101%
81-102%
90-98%
EDTA Plasma (n = 5)
83-96%
84-101%
84-101%
Heparin Plasma (n = 5)
82-98%
87-101%
82-99%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 5)
93-102
96
EDTA Plasma (n = 5)
88-99
95
Heparin Plasma (n = 5)
89-101
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 reagents and TMB substrate to room temperature. Set standard, test sample and control (zero) wells on the pre-coated plate and record their positions.
2
Primary Incubation: Prepare standards, samples, blanks and load into designated wells. Incubate plate at 37°C for 90 minutes to allow antigen binding.
3
Detection Antibody Binding: Add biotin-labeled detection antibody and incubate at 37°C for 60 minutes.
4
HRP-Streptavidin Binding: Add HRP-Streptavidin (SABC) and incubate at 37°C for 30 minutes.
5
Color Development: Add TMB substrate and incubate in the dark for 10–20 minutes.
6
Stop Reaction & Reading: Add stop solution and measure absorbance at 450 nm immediately.
Sample Type
Protocol
Serum
Allow blood to clot, centrifuge at 1000 × g for 20 minutes, collect supernatant supernatant and store appropriately.
Plasma
Collect using anticoagulant tubes, centrifuge at 1000 × g for 15 minutes at 2–8°C and collect plasma.
Tissue Homogenate
Homogenize tissue in PBS with protease inhibitors, centrifuge and collect supernatant.
Cell Culture Supernatant
Centrifuge at 2500 rpm for 5 minutes and collect clarified supernatant.
Cell Lysate
Lyse cells using lysis buffer with protease inhibitors, centrifuge and collect protein supernatant.
Other Sample Types
For more information about how to process other sample types, (e.g., body fluids, breast milk & more), please contact our Tech Support Team at 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.
