Rat TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit

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This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti TNF-α antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with TNF-α bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of TNF-α in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Catalogue No.: 
ER1393
Alias: 
TNF-α ELISA Kit, DIF ELISA Kit, TNF-alpha ELISA Kit, TNFA ELISA Kit, TNFSF2 ELISA Kit
Species: 
Rat
Range: 
3.906-250pg/ml
Sensitivity: 
2.344pg/ml
Manual MSDS
  • SPECIFICATIONS
  • CITATIONS
Product Name
Rat TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit
Alias
TNF-α ELISA Kit, DIF ELISA Kit, TNF-alpha ELISA Kit, TNFA ELISA Kit, TNFSF2 ELISA Kit
Catalogue No.
ER1393
Size
48T/96T
Species
Rat
UniProt No.
P16599
Sample Type
Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method
Sandwich ELISA, Double Antibody
Detection Wavelength
OD450
Reaction Duration
4 hours
Range
3.906-250pg/ml
Sensitivity
2.344pg/ml
Storage
2-8°C(Sealed), Don't cryopreserve.
Specificity
Specifically binds with TNF-α , no obvious cross reaction with other analogues.
ELISA Kit Components
Kit Components Item Size(48T) Size(96T) Storage Condition for Opened Kit
E001 ELISA Microplate(Dismountable) 8×6 8×12 Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002 Lyophilized Standard 1vial 2vial Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003 Biotin-labeled Antibody(Concentrated, 100X) 60ul 120ul 2-8°C (Avoid Direct Light)
E034 HRP-Streptavidin Conjugate(SABC, 100X) 60ul 120ul
E024 TMB Substrate 5ml 10ml
E039 Sample Dilution Buffer 10ml 20ml 2-8°C
E040 Antibody Dilution Buffer 5ml 10ml
E049 SABC Dilution Buffer 5ml 10ml
E026 Stop Solution 5ml 10ml
E038 Wash Buffer(Concentrated, 25X) 15ml 30ml
E006 Plate Sealer 3 pieces 5 pieces  
E007 Product Description 1 copy 1 copy
Required Instruments and Reagents
  1. Microplate reader (wavelength: 450nm)
  2. 37°C incubator (CO2 incubator for cell culture is not recommenced.)
  3. Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
  4. Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
  5. Sterile tubes and Eppendorf tubes with disposable tips
  6. Absorbent paper and loading slot
  7. Deionized or distilled water
Assay Procedure Summary
Elisa实验原理图
  • Step 1: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
  • Washing: Wash the plate twice without immersing.
  • Step 2: Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37°C.
  • Washing: Wash the plate three times and immerse for 1min each time.
  • Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37°C.
  • Washing: Wash the plate five times and immerse for 1min each time.
  • Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
  • Step 5: Add 50ul stop solution. Read at 450nm immediately and calculate.
Standard Curve

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)

Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(pg/ml) OD-1 OD-2 Average
0 0.088 0.092 0.09
3.906 0.111 0.116 0.113
7.813 0.134 0.14 0.137
15.625 0.264 0.274 0.269
31.25 0.433 0.45 0.442
62.5 0.742 0.772 0.757
125 1.242 1.292 1.267
250 2.005 2.087 2.046
ER1393 Standard Curve Image
Recovery

Add a certain amount of TNF-α into the sample. Calculate the recovery by comparing the measured value with the expected amount of TNF-α in the sample.

Sample Type Recovery Range(%) Average(%)
serum(n=10) 88-96 93
EDTA plasma(n=10) 90-97 94
Heparin plasma(n=10) 86-104 96
Linearity

Dilute the sample with a certain amount of TNF-α at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type 1:2 1:4 1:8
serum(n=10) 89-93% 86-105% 87-100%
EDTA plasma(n=10) 85-98% 82-96% 83-101%
Heparin plasma(n=10) 81-100% 81-99% 84-100%
Precision(%)

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.

Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/ml) 7.78 30.39 124.29 8.11 30.97 125.2
Standard deviation 0.48 1.34 5.56 0.36 1.46 6
CV(%) 6.2 4.42 4.47 4.5 4.72 4.79
Stability

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5) 37°C for 1 month 2-8°C for 6 months
Average(%) 80 95-100
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2020-09-16
Product:
Protective Effect of S-Allyl Cysteine Against Neonatal Asthmatic Rats
Journal:
research square
Cited Date:
2020-09-29
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Basil (Ocimum basilicum) Leaves Essential Oil Ameliorates GluR1 Receptor Expression, TNF-α Level, and Pain-like Behaviors in Post-operative Pain Setting
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Bali Journal of Anesthesiology
Cited Date:
2022-05-27
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Chitosan/Alginate-loaded Astragalus hamosus shows ameliorating effects on lipid profile, inflammatory and hormonal parameters, and reduces miRNA-222 expression in Polycystic Ovary Syndrome Rats
Journal:
Research Square
Author:
Islamic Azad University
Cited Date:
2024-02-18
Product:
Evaluating the effect of hydro-alcoholic extract of Phoenix dactylifera L. spathe on streptozotocin-induced diabetic rats
Journal:
Comparative Clinical Pathology
Cited Date:
2021-06-25
Product:
Antidiabetic, Antioxidant and Anti-Inflammatory Activities of Residual Aqueous Fraction of Ethulia conyzoides in Induced Type 2 Diabetic Rats
Journal:
Tropical Life Sciences Research
Cited Date:
2022-11-18
Product:
Assessment of the Therapeutic Potential of a Novel Phosphoramidate Acyclic Nucleosideon Induced Hepatocellular Carcinoma in Rat Model
Journal:
SSRN
Author:
Biotechnology Program, Basic and Applied Sciences Institute, Egypt-Japan University of Science and Technology (E-JUST), 21934, New Borg El-Arab City, Alexandria, Egypt
Cited Date:
2024-03-01
Product:
Potential effects of incretin-based therapies on polycystic ovary syndrome in rats: a comparative study of linagliptin versus liraglutide
Journal:
International Journal of Research in Pharmaceutical Sciences
Cited Date:
2021-07-15
Product:
Synergistic effect of oral zinc and dietary M. oleifera Leaf Supplementation on Diabetes-induced Oxidative Stress and Inflammation
Journal:
Sahel Journal of Life Sciences FUDMA
Author:
Department of Biochemistry and Molecular Biology, Federal University Dutsin-Ma, P.M.B 5001, Katsina State, Nigeria
Sample:
serum
Cited Date:
2024-03-08
Product:
Phytosterols content of Keunnunjami germ and its antioxidative effects in adult rats
Journal:
Journal of Nutrition and Health
Cited Date:
2020-03-13
Product:
Therapeutic potential of oxymatrine in impeding the cardiomyopathy in the STZ-nicotinamide induced diabetes through SIRT1/Nrf2 signaling activation
Journal:
Research Square
Cited Date:
2022-12-09
Product:
Gallic acid reduces inflammatory cytokines and markers of oxidative damage in a rat model of estradiol-induced polycystic ovary
Journal:
Comparative Clinical Pathology
Cited Date:
2019-02-15
Product:
Sinapic Acid Ameliorates Cisplatin-Induced Peripheral Neuropathy: An In Vivo and In Vitro Study
Journal:
F?rat University Medical Journal of Health Sciences
Author:
Gaziantep University, Faculty of Medicine, Department of Physiology, Gaziantep, T?RK?YE
Cited Date:
2023-07-07
Product:
Zinc Supplementation in Cytokine Regulation During LPS-induced Sepsis in Rodent
Journal:
Journal of International Dental and Medical Research
Cited Date:
2020-05-02
Product:
Selenium Attenuates Cholestasis-Induced Liver Injury and Fibrosis by Alleviating Liver Oxidative Stress and Inflammation in Rats
Journal:
Bulletin of Egyptian Society for Physiological Sciences
Cited Date:
2022-03-10
Product:
Effects of Hydroalcoholic Leaf Extract of Avicennia marina on Apoptotic, Inflammatory, Oxidative Stress, and Lipid Peroxidation Indices and Liver Histology of Type 1 Diabetic Rats
Journal:
HEPATITIS MONTHLY
Cited Date:
2020-05-10
Product:
Vanillic acid attenuates testicular ischemia/reperfusion injury in rats
Journal:
Beni-Suef University Journal of Basic and Applied Sciences
Cited Date:
2023-01-06
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