The Porcine TNF-alpha ELISA Kit is a highly sensitive and specific assay designed for the accurate detection of TNF-alpha levels in porcine samples, including serum, plasma, and cell culture supernatants. This kit provides reliable and reproducible results, making it ideal for a variety of research applications. TNF-alpha is a key cytokine involved in inflammation, immune response, and cell death. Dysregulation of TNF-alpha levels is associated with various diseases, including autoimmune disorders, inflammatory conditions, and infectious diseases.
Monitoring TNF-alpha levels in porcine samples can provide valuable insights into the mechanisms underlying these diseases and aid in the development of potential therapeutic strategies. Overall, the Porcine TNF-alpha ELISA Kit is a valuable tool for researchers studying the role of TNF-alpha in porcine health and disease, offering accurate and precise measurement of this important cytokine in biological samples.
Product Name:
Porcine TNF alpha ELISA Kit
SKU:
PRFI00158
Reactivity:
Porcine
Assay Type:
Sandwich ELISA, Double Antibody
Sensitivity:
18.75 pg/mL
Range:
31.25-2000 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)
85-101%
83-99%
85-97%
EDTA Plasma (n = 5)
84-99%
85-101%
84-95%
Heparin Plasma (n = 5)
84-99%
90-104%
84-99%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 5)
93-99
98
EDTA Plasma (n = 5)
86-99
96
Heparin Plasma (n = 5)
88-105
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 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
-
Technical Manual
1 copy
1 copy
-
Browne et al.
Yeast cell wall products effect on attachment of harmful foodborne strains of Salmonella to intestinal porcine enterocyte cell line (IPEC-J2)
