The Canine TNF-alpha ELISA Kit is specially developed for the precise measurement of TNF-alpha levels in canine serum, plasma, and cell culture supernatants. With its superior sensitivity and specificity, this kit delivers consistent and accurate results, making it an invaluable tool for various research purposes. TNF-alpha is a key pro-inflammatory cytokine that plays a crucial role in the immune response, inflammation, and various disease processes in dogs.
By accurately quantifying TNF-alpha levels, researchers can gain insights into the underlying mechanisms of inflammatory conditions, infectious diseases, and autoimmune disorders in canines. Overall, the Canine TNF-alpha ELISA Kit is essential for studying the role of TNF-alpha in canine health and disease, as well as for developing potential therapeutic interventions to improve the well-being of man's best friend.
Product Name:
Canine TNF alpha ELISA Kit
SKU:
CNFI00020
Reactivity:
Canine
Assay Type:
Sandwich ELISA, Double Antibody
Sensitivity:
9.375 pg/mL
Range:
15.625-1000 pg/mL
Sample Type:
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)
87-104%
97-103%
86-103%
EDTA Plasma (n = 5)
83-98%
85-101%
93-101%
Heparin Plasma (n = 5)
82-99%
84-99%
82-100%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 5)
86-100
94
EDTA Plasma (n = 5)
90-105
98
Heparin Plasma (n = 5)
88-96
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 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.
Biotin-labeled Antibody (Concentrated, 100X)
60 ul
120 ul
2-8°C (Avoid direct light)
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)
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
10 ml
2-8°C
Wash Buffer(25X)
15 ml
30 ml
2-8°C
Plate Sealer
3 pieces
5 pieces
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Technical Manual
1 copy
1 copy
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