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: 
Tumor necrosis factor ELISA Kit, Cachectin ELISA Kit, TNF-alpha ELISA Kit, Tumor necrosis factor ligand superfamily member 2 ELISA Kit, TNF-a ELISA Kit, Tumor necrosis factor, membrane form ELISA Kit, N-terminal fragment ELISA Kit, NTF ELISA Kit, Intracellular domain 1 ELISA Kit, ICD1 ELISA Kit, Intracellular domain 2 ELISA Kit, ICD2 ELISA Kit, C-domain 1 ELISA Kit, C-domain 2 ELISA Kit, Tumor necrosis factor, soluble form ELISA Kit, TNF ELISA Kit, TNFA ELISA Kit, TNFSF2 ELISA Kit
Species: 
Rat
Range: 
3.906-250pg/ml
Sensitivity: 
2.344pg/ml
Manual MSDS
  • SPECIFICATIONS
  • CITATIONS
  • FIGURES
Product Name
Rat TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit
Alias
Tumor necrosis factor ELISA Kit, Cachectin ELISA Kit, TNF-alpha ELISA Kit, Tumor necrosis factor ligand superfamily member 2 ELISA Kit, TNF-a ELISA Kit, Tumor necrosis factor, membrane form ELISA Kit, N-terminal fragment ELISA Kit, NTF ELISA Kit, Intracellular domain 1 ELISA Kit, ICD1 ELISA Kit, Intracellular domain 2 ELISA Kit, ICD2 ELISA Kit, C-domain 1 ELISA Kit, C-domain 2 ELISA Kit, Tumor necrosis factor, soluble form ELISA Kit, TNF 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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Anti-inflammatory activity of Turkey source pumpkin seed oil in rat oedema model
IF: 0.877
Journal:
Acta Poloniae Pharmaceutica - Drug Research
Cited Date:
2020-03-13
Product:
Ethanolic extracts of babandotan leaves (Ageratum conyzoides L.) prevents inflammation and proteoglycan degradation by inhibiting TNF-α and MMP-9 on osteoarthritis rats induced by monosodium iodoacetate
IF: 0.841
Journal:
Asian Pacific Journal of Tropical Medicine
Cited Date:
2017-03-06
Product:
Gallic acid attenuates torsion/detorsion-induced testicular injury in rats through suppressing of HMGB1/NF-κB axis and endoplasmic reticulum stress
IF: 0.8
Journal:
Revista Internacional de Andrología
Author:
Department of Nutrition and Dietetics, Faculty of Health Sciences, Karadeniz Technical University, 61080 Trabzon, Turkey
Cited Date:
2024-05-06
Product:
Tualang honey and DHA-rich fish oil reduce the production of pro-inflammatory cytokines in the rat brain following exposure to chronic stress
IF: 0.78
Journal:
Journal of Taibah University Medical Sciences
Cited Date:
2019-07-23
Product:
Hesperidin enhances angiogenesis via modulating expression of growth and inflammatory factor in diabetic foot ulcer in rats
IF: 0.719
Journal:
European Journal of Inflammation
Cited Date:
2018-05-14
Product:
Protocatechuic acid attenuates cerebral aneurysm formation and progression by inhibiting TNF-alpha/Nrf-2/NF-kB-mediated inflammatory mechanisms in experimental rats
IF: 0.69
Journal:
Open Life Sciences
Cited Date:
2021-02-26
Product:
High Fiber Diet Suppression of TLR4 and NFκβ Gene Expression Correlated with TNF-α and IL-6 Levels in Hypertriglyceridemia Rats
IF: 0.687
Journal:
Journal of Food and Nutrition Research
Cited Date:
2018-12-19
Product:
Protective effect of S-allyl cysteine against cerebral ischemia/reperfusion injury in experimental rats
IF: 0.533
Journal:
Tropical Journal of Pharmaceutical Research
Cited Date:
2021-10-08
Product:
Simvastatin suppresses cerebral aneurysm in rats through suppression of release of pro-inflammatory cytokines
IF: 0.533
Journal:
Tropical Journal of Pharmaceutical Research
Cited Date:
2022-06-17
Product:
The effect of crocin on apoptotic, inflammatory, BDNF, Pt, and Aβ40 indicators and neuronal density of CA1, CA2, and CA3 regions of hippocampus in the model of Alzheimer suffering rats induced with trimethyltin chloride
Journal:
Comparative Clinical Pathology
Cited Date:
2019-06-25
Product:
Hepatocurative Effects of Alpha Lipoic Acid in 5-Fluorouracil-Induced Toxicity Involves Activation of TNF Alpha and NF-κβ Signaling Pathway
Journal:
Tropical Journal of Natural Product Research
Cited Date:
2021-02-26
Product:
Anti-Inflammatory Effect of Omega-3 Fish Oil Accelerates MyoD and Myogenin Expressions Following Rat Exercise-Induced Muscle Injury Model in a Dose-Dependent Manner
Journal:
Malaysian Journal of Medicine and Health Sciences (MJMHS)
Cited Date:
2020-06-30
Product:
Crataegus sinaica defatted methanolic extract ameliorated monosodium iodoacetate-induced oxidative stress andinhibited inflammation in a rat model of osteoarthritis
Journal:
Research in Pharmaceutical Sciences
Cited Date:
2022-09-08
Product:
MEDICINAL COMPOSITION OF AMARANTH ORIGIN FOR CARDIOVASCULAR TREATMENT
Journal:
United States Patent Application
Cited Date:
2020-07-16
Product:
Toxicological implications of sequential administration of herbal and conventional medicines: Evidence from an in vivo study on Azadirachta indica and artesunate in male Wistar rats
Journal:
Toxicology Research and Application
Cited Date:
2021-03-25
Product:
Improvement of Pharmaceutically Active Materials from the Fruit of the Prickly Pear (Opuntia ficus-indica) Using Ultrasound (UAE) and Microwave (MAE) Assisted Extractions
Journal:
Biointerface Research in Applied Chemistry
Cited Date:
2023-02-09
Product:
Application of Platelet-Rich Plasma in Severe Acute Experimental Pancreatitis
Journal:
Novosti Khirurgii
Author:
Department of Surgery and Transplantation, Belarusian State Medical University, Minsk, Dzerzhinsky Ave. 83, 220116, Republic of Belarus
Cited Date:
2023-10-08
Product:
Antagonistic effectiveness of Anacardium occidentale leaf extract on lead-acetate exposure-induced hepatorenal toxicity in rats
Journal:
Environmental Analysis Health and Toxicology
Author:
Department of Human Anatomy, Kaduna State University, State, Nigeria
Cited Date:
2024-01-19
Product:
Mesenchymal Stem Cell Transplantation Promotes Functional Recovery through MMP2/STAT3 Related Astrogliosis after Spinal Cord Injury
Journal:
International Journal of Stem Cells
Cited Date:
2019-07-30
Product:
Regulatory Effect of Secretome-Hypoxia Mesenchymal Stem Cells on TNF-α Level in Streptozotocin-induced Rats
Journal:
International Journal of Cell and Biomedical Science
Cited Date:
2023-02-10
Product:
The Effect of Sapodilla Leaf Extract (Manilkara Zapota LP Royen) on IL-1 and TNF-α Expression in Male White Rats Wistar Strain Induced with UVB Light
Journal:
International Journal of Multidisciplinary Research and Analysis
Author:
Faculty of Medicine, Universitas Islam Sultan Agung, Jl Kaligawe KM 4 Semarang 50012
Cited Date:
2024-05-06
Product:
N-Acetylcysteine Ameliorates 5-Fluorouracil‐Induced Ovarian Injury in Rats
Journal:
Clinical and Experimental Health Sciences
Author:
KARADENIZ TECHNICAL UNIVERSITY
Cited Date:
2023-10-20
Product:
The use of ultrasound (UAE) and microwaves (MAE) to improve the extraction of pharmaceutically active materials from the fruit of the prickly pear (Opuntia ficus-indica)
Journal:
Research Square
Cited Date:
2022-05-05
Product:
The effect of PRP in oxidative stress male rats induced by D-galactose
Journal:
AIP Conference Proceedings
Cited Date:
2020-09-16
Product:
Protective Effect of S-Allyl Cysteine Against Neonatal Asthmatic Rats
Journal:
research square
Cited Date:
2020-09-29
Product:
Basil (Ocimum basilicum) Leaves Essential Oil Ameliorates GluR1 Receptor Expression, TNF-α Level, and Pain-like Behaviors in Post-operative Pain Setting
Journal:
Bali Journal of Anesthesiology
Cited Date:
2022-05-27
Product:
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:
Downregulation of P2y14 Receptor Expression Ameliorates Diabetic Cardiac Autonomic Neuropathy by Inhibiting Glial Ferroptosis Via the Nrf2/Gpx4 Axis
Journal:
SSRN
Author:
School of Basic Medical Sciences, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi 330006, P.R. China
Sample:
SCG and neuron culture medium
Cited Date:
2024-07-26
Product:
Effect of Omega-3 Fatty Acid on Osteoanabolic Markers and Bone-resorbing Cytokines in Ovariectomized Rats
Journal:
Journal of Innovative Research in Life Sciences
Author:
Department of Biochemistry, Federal University Gusau, Zamfara State-Nigeria
Sample:
serum
Cited Date:
2024-05-31
Product:
The Modulatory Role of L-carnitine on Inflammation in Experimentally-induced Testicular Torsion in Wistar Rats
Journal:
FUDMA Journal of Sciences
Author:
Department of Human Physiology, Faculty of Basic Medical Science, Ahmadu Bello University, Zaria, Nigeria.
Sample:
serum
Cited Date:
2024-08-02
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:
Therapeutic Effect of Astaxanthin on 5-Fluorouracil-Induced Ovarian Damage in Rats
Journal:
Bilecik Seyh Edebali University Journal of Science
Author:
Department of Chemistry and Chemical Processing Technologies, Macka Vocational School, Karadeniz Technical University, 61750 Trabzon, Türkiye
Sample:
supernatants
Cited Date:
2024-06-07
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:
Efficacy of nanoencapsulated Moringa oleifera L. seeds and Ocimum tenuiflorum L. leaves extracts incorporated in functional soft cheese on streptozotocin-induced diabetic rats
Journal:
Phytomedicine Plus
Author:
Dairy Department, Food Industries and Nutrition Research Institute, National Research Centre, Cairo 12622, Egypt
Species:
blood samples
Cited Date:
2024-06-14
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
Product:
Indole-3-propionic acid alleviates chondrocytes inflammation and osteoarthritis via the AhR/NF-κB axis.
Molecular medicine (Cambridge, Mass.)
2023-1-31
Indole-3-propionic acid alleviates chondrocytes inflammation and osteoarthritis via the AhR/NF-κB axis.
Huangming Zhuang, Xunshan Ren, Fuze Jiang, Panghu Zhou
MiR-495-3p depletion contributes to myocardial ischemia/reperfusion injury in cardiomyocytes by targeting TNC.
Regenerative therapy
2022-12
MiR-495-3p depletion contributes to myocardial ischemia/reperfusion injury in cardiomyocytes by targeting TNC.
Wei Song, Naiyan Qiu
Supported by deepbiogroup
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