Mouse Tumor necrosis factor α,TNF-α ELISA KIT

Code CSB-E04741m
Size 96T,5×96T,10×96T
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Trial Size 24T ELISA Kit Trial Size (Only USD$150/ kit)
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Product Details

Alternative Names
Tnf ELISA Kit; Tnfa ELISA Kit; Tnfsf2 ELISA Kit; Tumor necrosis factor ELISA Kit; Cachectin ELISA Kit; TNF-alpha ELISA Kit; Tumor necrosis factor ligand superfamily member 2 ELISA Kit; TNF-a) [Cleaved into: Tumor necrosis factor ELISA Kit; membrane form ELISA Kit; N-terminal fragment ELISA Kit; NTF); Intracellular domain 1 ELISA Kit; ICD1); Intracellular domain 2 ELISA Kit; ICD2); C-domain 1; C-domain 2; Tumor necrosis factor ELISA Kit; soluble form] ELISA Kit
Abbreviation
Uniprot No.
Species
Mus musculus (Mouse)
Sample Types
serum, plasma, cell culture supernates, tissue homogenates
Detection Range
15.6 pg/ml-1000 pg/ml
Sensitivity
3.9 pg/ml
Assay Time
1-5h
Sample Volume
50-100ul
Detection Wavelength
450 nm
Research Area
Immunology
Assay Principle
quantitative
Measurement
Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%  
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%  
Three samples of known concentration were tested in twenty assays to assess.
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of mouse TNF-α in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
 
  Sample Serum(n=4)  
1:1 Average % 86  
  Range % 80-92  
1:2 Average % 95  
  Range % 91-105  
1:4 Average % 100  
  Range % 92-110  
1:8 Average % 92  
  Range % 86-98
 
Recovery
The recovery of mouse TNF-α spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
 
Sample Type Average % Recovery Range  
Serum (n=5)   93 89-98  
EDTA plasma (n=4) 95 90-100
 
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/ml OD1 OD2 Average Corrected  
1000 2.793 2.822 2.808 2.651  
500 1.889 1.943 1.916 1.759  
250 1.245 1.211 1.228 1.071  
125 0.729 0.686 0.708 0.551  
62.5 0.373 0.404 0.388 0.232  
31.2 0.287 0.298 0.292 0.136  
15.6 0.231 0.244 0.238 0.081  
0 0.152 0.161 0.157  
 
Materials provided
    • An assay plate --- The 96-well plate is composed of 12 x 8 strip plates and can be taken apart.
    • Two bottles of lyophilized standard --- Reconstitute one bottle of standard with sample diluent at dilution series and draw the standard curve.
    • Biotin-labeled TNF-α antibody (100 x concentrate) 1 x 120 μl --- Act as the detection antibody and need to be diluted before use.
    • HRP-avidin (100 x concentrate) 1 x 120 μl --- Catalyze the TMB substrate for color development and need to be diluted before use.
    • Biotin-antibody Diluent 1 x 15 ml --- Dilute the Biotin-antibody.
    • HRP-avidin Diluent 1 x 15 ml --- Dilute the HRP-avidin.
    • Sample Diluent 1 x 50 ml --- Reconstitute the standard and dilute the sample to an appropriate concentration.
    • Wash Buffer (25 x concentrate) 1 x 20 ml --- Wash away unbound solution and non-specific substances.
    • TMB Substrate 1 x 10 ml --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
    • Stop Solution 1 x 10 ml --- Add stop solution and stop the color development. The solution color immediately turns from blue to yellow.
    • Four Adhesive Strips (For 96 wells) --- Seal the microtiter plate when incubation.
    • An instruction manual
Materials not provided
    • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
    • An incubator can provide stable incubation conditions up to 37°C±5°C.
    • Centrifuge
    • Vortex
    • Squirt bottle, manifold dispenser, or automated microplate washer
    • Absorbent paper for blotting the microtiter plate
    • 50-300ul multi-channel micropipette
    • Pipette tips
    • Single-channel micropipette with different ranges
    • 100ml and 500ml graduated cylinders
    • Deionized or distilled water
    • Timer
    • Test tubes for dilution
Troubleshooting
and FAQs
Storage
Store at 2-8°C. Please refer to protocol.
Shelf Life
6 months
Lead Time
3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx.
Description

This Mouse TNF ELISA Kit was designed for the quantitative measurement of Mouse TNF protein in serum, plasma, cell culture supernates, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 15.6 pg/mL-1000 pg/mL and the sensitivity is 3.9 pg/mL.

Citations

Customer Reviews and Q&A

 Customer Reviews
Average Rating:
5.0 - 6 reviews

Submit a Review here

Sample type: Cell culture supernatant

Sample species: Mouse

Sample dilution: No dilution

Review: I used CSB-E04741m to detect TNFa levels in cell supernatant exosomes and mouse supernatants, respectively. The results of the standard curve were good.

By Anonymous

Sample type: Cell culture supernatant

Sample species: Mouse

Sample dilution: No dilution

Review: I used the CSB-E04741m kit to detect the levels of TNF-a in the homogenized hippocampal tissue samples of mice before and after modeling as well as before and after treatment. The standard curve results were excellent, and it was found that TNF-a levels increased after modeling while they decreased after treatment, which is consistent with the expected results. The kit was convenient to use with simple and efficient steps, allowing for saved experimental time.

By Anonymous

Sample type: Tissue homogenate

Sample species: Mouse

Sample dilution: No dilution

Review: I have tested the CSB-E04741m for PR8 virus-infected mouse lung tissue homogenates, the operation is simple and easy to use, the sensitivity is very high, and the test results are very accurate.

By Anonymous

Sample type: Serum

Sample species: Mouse

Sample dilution: 1: 5

Review: I used CSB-E04741m to detect spontaneous T2DM mice db/db mice. The dilution ratio was 1: 5, the dilution ratio was appropriate and the concentration was moderate. The measured results were accurate and cost-effective!

By Anonymous

Sample type: Serum

Sample species: Mouse

Sample dilution: No dilution

Review: The linear correlation of calibration curve is good > 0.9999, and the sample concentration is located in the second half of calibration curve.

By Anonymous

Sample type: Serum

Sample species: Mouse

Sample dilution: No dilution

Review: I have detected influenza virus infection in the peripheral blood supernatant of mice using CSB-E04741m. The operation is simple, convenient, and easy to get started, with very high sensitivity and very accurate detection results.

By Anonymous

 Q&A
Q:

I need a Elisa analysis of TNF alpha on macrophase cell and lymphocytes

A:
Thanks for your inquiry.
Regarding elisa kit for mouse TNF-α,we have the following kit for you:
#CSB-E04741m,Mouse Tumor necrosis factor α,TNF-α ELISA KIT
This kit is suitable to detect cell culture supernates. I will send you the kit manual by email.
But for macrophase cell and lymphocytes, sorry we did not test either of them with this kit before.We suggest you do pre-testing if you want to test macrophase cell and lymphocytes.

Target Background

Function
(From Uniprot)
Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. Induces insulin resistance in adipocytes via inhibition of insulin-induced IRS1 tyrosine phosphorylation and insulin-induced glucose uptake. Induces GKAP42 protein degradation in adipocytes which is partially responsible for TNF-induced insulin resistance. Plays a role in angiogenesis by inducing VEGF production synergistically with IL1B and IL6.; The TNF intracellular domain (ICD) form induces IL12 production in dendritic cells.
Gene References into Functions
  1. Following Unfolded Protein Response activation during autosomal dominant retinitis pigmentosa progression, secrete TNFalpha and signal a self-destructive program to the cones, resulting in their cell death. PMID: 27750040
  2. In a hypercaloric environment, persistent elevation of microglial reactivity and consequent TNFalpha secretion induces mitochondrial stress in POMC neurons that contributes to the development of obesity. Specific disruption of the gene expressions of TNFalpha downstream signals TNFSF11A or NDUFAB1 in the mediobasal hypothalamus of diet-induced obese mice reverses mitochondrial elongation and reduces obesity. PMID: 28489068
  3. persistent stimulation with titanium particles may lead to a consistent release of TNF-alpha and IL-6 via SPHK-2 activity, which may lead to aseptic implant loosening PMID: 29728804
  4. Recognition memory improved with exercise in WT mice, was impaired in TNFR1(-/-) exercise mice, showed non-significant impairment with exercise in TNF(-/-) mice, and no changes in TNFR2(-/-) mice. In spatial learning there were exercise related improvements in WT mice, non-significant but meaningful impairments evident in TNFR1(-/-) exercise mice, modest improvement in TNF(-/-) exercise mice. PMID: 29969604
  5. In vitro mild uncoupling rescued from TNF-induced endothelial permeability, disassembly of cell contacts and VE-cadherin cleavage by the matrix metalloprotease 9 (capital EM, Cyrilliccapital EM, Cyrilliccapital ER, Cyrillic9). The uncouplers prevented TNF-induced expression of MMP9 via inhibition of NFkappaB signaling. PMID: 28131916
  6. macrophage-TNF-induced AKT/beta-catenin signalling in Lgr5(+) hair follicle stem cells has a crucial role in promoting hair follicle cycling and neogenesis after wounding PMID: 28345588
  7. Transmembrane TNF, TNFR2 and TNFR1 (indirectly) are critical for preventing inflammation during BCG-induced pleurisy in mice. PMID: 29973541
  8. findings demonstrate a new role for TNFalpha as a key regulator of neutrophil trafficking into and within lymphatic system in vivo. PMID: 28287124
  9. Our work suggested that TNF-alpha and TNF-R1 are the major contributors of TNF signaling pathway in anesthesia-induced spinal cord neurotoxicity. Targeting TNF-alpha / TNF-R1, not TNF-R2 signaling pathway may be the key component to rescue or prevent anesthesia-induced apoptotic injury in spinal cord neurons. PMID: 29802833
  10. observation from the present research work reveals that Quercetin suppressed the production of proinflammatory cytokines at different levels, such as TNF-alpha and IL-1beta, and inhibits the activation of I-kappaB phosphorylation, whereas the total content was not affected. PMID: 29322353
  11. this is the first evidence to suggest that TET2 mutations promote clonal dominance with aging by conferring TNFalpha resistance to sensitive bone marrow progenitors while also propagating such an inflammatory environment. PMID: 29195897
  12. Elevated A20 promotes TNF-induced and RIPK1-dependent intestinal epithelial cell death PMID: 30209212
  13. M. tuberculosis and TNFalpha synergise to induce necroptosis in murine fibroblasts via RIPK1-dependent mechanisms and characterized by phosphorylation of Ser345 of the MLKL necroptosis death effector. PMID: 28892415
  14. Our current study has demonstrated that in allergic airway disease (AAD) mice, intestinal dysbiosis (ID) caused increased nasal rubbing, sneezing, serum OVA specific IgE level and pro-inflammatory cytokine TNF-alpha in NALF and BALF. ID also inhibited miR-130a expression in AAD mice. Further molecular experiments indicated that miR-130a could specifically target and repress TNF-alpha mRNA expression. PMID: 29702281
  15. These data may indicate that insulin resistance in Adp(-/-) mice is likely caused by an increase in concentrations of TNFalpha and FFA via downregulation of PPARalpha. PMID: 29445073
  16. TNF-alpha is involved in cardiac PHLPP1 upregulation during reoxygenation, which is mediated by NF-kappaB transcriptional activity PMID: 29940243
  17. Lack of TNF-alpha signaling through Tnfr1 makes the mice more susceptible to acute infection but does not alter state of latency and reactivation of HSV-1. PMID: 29113822
  18. although TNFalpha does not induce osteoclastogenesis alone, it does work with RANKL to induce osteoclastic differentiation, and the NFkappaB pathway may serve an important role in this process. PMID: 29512766
  19. two different modes of necroptosis induction by TNFalpha exist which are differentially regulated by iuRIPK1 formation. Overall, this work reveals a distinct mechanism of RIPK1 activation that mediates the signaling mechanism of RDA as well as a type of necroptosis. PMID: 29891719
  20. Results demonstrate a critical role for the TRPM2 channel in Abeta42-induced microglial activation and generation of TNF-alpha: PKC/NOX-mediated generation of ROS and activation of PARP-1 are required for Abeta42-induced TRPM2 channel activation and, furthermore, the PYK2/MEK/ERK signaling pathway as a positive feedback mechanism downstream of TRPM2 channel activation facilitates further activation of PARP-1 and TRPM2 ... PMID: 29143372
  21. TNFalpha may act reciprocally with DRA, leading to the development of intestinal inflammation. PMID: 29286110
  22. TNF-alpha plays a pivotal role in the development of nonalcoholic fatty liver disease and progression to nonalcoholic steatohepatitis. PMID: 28922680
  23. Cross-fostering and conditional knockout experiments indicated that a TNF-alpha deficit in the maternal brain, rather than in the hematopoietic system, and during gestation was responsible for the low-fear offspring phenotype. PMID: 29199072
  24. In a retinitis pigmentosa mouse model, TrkC activity generates phosphorylated Erk, which upregulates glial TNF-alpha, causing selective neuronal death. PMID: 29242588
  25. genome-wide knockdown of 19 ribosomal proteins resulted in decreased IL-10 and increased TNF-alpha production. PMID: 29657255
  26. We conclude that one of the possible regulatory mechanisms of TNF in mechanical orofacial hyperalgesia involves upregulation of the nociceptor TRPV1 PMID: 29132095
  27. The work highlighted the modulatory role of miR-105 in TNF-alpha-induced epithelial-mesenchymal transition and promoting colorectal cancer metastasis. PMID: 29238068
  28. These results suggest that glucocorticoids' effects on adipose tissue immune response, both in a pro- and an anti-inflammatory manner, depend on the nutritional status. PMID: 29847081
  29. This study demonstrated that TNF-alpha genetic deletion ameliorates the amyloid phenotype of the 5XFAD mouse model of AD. 5XFAD/TNF-alpha-/- mice exhibit significantly decreased amyloid deposition and reduced levels of AbetaPP-CTFs and amyloid-beta protein. PMID: 28826177
  30. Data suggest that expression of Tnfa in adipocytes can be regulated by dietary fatty acids; here, polyunsaturated fatty acids regulate Tnfa expression via alteration in methylation of Tnfa promoter in rats fed polyunsaturated fatty acids (safflower oil versus coconut/olive oil) and in mouse adipocyte cell line incubated with polyunsaturated fatty acid (linoleic acid versus palmitic/oleic acids). PMID: 28575756
  31. a precise mechanism for attenuation of HgCl2-induced liver dysfunction by dietary luteolin via regulating Sirt1/Nrf2/TNF-alpha signaling pathway, and provide a foundation for further study of luteolin as a novel therapeutic agent against inorganic mercury poisoning. PMID: 27853236
  32. a significant increase in plasma levels of IL-2, IFN-g and TNF-a was revealed as assessed by ELISA. In conclusion, the results of the present study indicate that MENK has a cytotoxic effect on B16 melanoma cells in vitro and in vivo, and suggest a potential mechanism for these bioactivities. PMID: 28849104
  33. findings suggest that PGRN deficiency leads to excessive NF-kappaB activation in microglia and elevated TNFalpha signaling, which in turn lead to hyperexcitability of medium spiny neurons and obsessive-compulsive behavior-like behavior. PMID: 28438992
  34. findings highlight an epigenetic mechanism by which EZH2 integrates the multifaceted effects of TNFalpha signaling to promote the inflammatory response and apoptosis in colitis. PMID: 28439030
  35. It is possible that JNK and TNF-alpha commonly contribute to kidney damage by assembling a positive feedback cycle after crush syndrome, leading to increased apoptosis in the renal cortex. HMGB1 from the muscle may be the trigger. PMID: 28701229
  36. Cytokine-inducing and anti-inflammatory activity of chitosan and its low-molecular derivative. PMID: 29513410
  37. Excessive death of hepatocytes is a characteristic of liver injury. A new programmed cell death pathway has been described involving upstream death ligands such as TNF and downstream kinases such as RIPK1. PMID: 28088582
  38. Taken together, we have demonstrated a role for TNF in the development of classically activated macrophages in listeriosis PMID: 28545808
  39. Inhibition of signaling stimulated by both TNF and IL1beta synergizes with NF-kappaB inhibition in eliminating leukemic stem cells. PMID: 28039479
  40. Calyptranthes grandifolia O.Berg (Myrtaceae) ethanolic extract inhibits TNF-alpha gene expression and cytokine release in vitro PMID: 28447740
  41. Results show that interleukin 6 (IL6) promotes oval cell proliferation and liver regeneration, while tumor necrosis factor alpha (TNFalpha) and TNF receptor-1(TNFR1) do not affect this process. PMID: 27556180
  42. This study adds to the evidence that both peripheral and brain region-specific increases in tumor necrosis factor alpha lead to both sickness and depression- and anxiety disorder-relevant behavior and do so via different pathways. PMID: 27515532
  43. Lactosylceramide-Induced Phosphorylation Signaling to Group IVA Phospholipase A2 via Reactive Oxygen Species in Tumor Necrosis Factor-alpha-Treated Cells. PMID: 28444900
  44. The current study demonstrated that honey can stimulate or suppress the mRNA expression of some pro-inflammatory cytokines in mice brains. Furthermore, honey suppresses the TNF-alpha mRNA expression in the presence of T. gondii infection but it stimulates the IL-1beta and IL-6 mRNA expression. Treatment of the mice with honey reduces parasite multiplication in the brain. PMID: 27591508
  45. aerobic interval training enhanced the anti-inflammatory indices IL-10/TNF-alpha ratio and IL-15 expression in skeletal muscle in tumor-bearing mice. PMID: 27863332
  46. findings suggest that activation of Tnf-Aicda axis and co-inhibitory signals to T cells in coordination with Th1-type immunity has critical roles in the immune response against Hepatitis B virus infection PMID: 28063995
  47. Taken together, we speculate that DT-13 inhibits endothelium vascular inflammation through regulating nitric oxide production and the expression of ROS, TNFR, IL-8, MCP-1, which are associated with inflammation. PMID: 29162452
  48. TNF signalling is required for the expansion and differentiation of pathogenic IFNgamma+CD4+ T cells that promote the irreversible loss of bone marrow function. PMID: 28671989
  49. Drugs targeting XIAP and cIAP1/2 may be effective for osteosarcoma patients whose tumors express abundant RIPK1 and contain high levels of TNFalpha. PMID: 27129149
  50. Taken together, we indicated that anti-IL-6 and anti-TNF-alpha therapy prevent intestinal permeability induced by intestinal inflammation PMID: 27155817

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Subcellular Location
Cell membrane; Single-pass type II membrane protein.; [Tumor necrosis factor, membrane form]: Membrane; Single-pass type II membrane protein.; [Tumor necrosis factor, soluble form]: Secreted.; [C-domain 1]: Secreted.; [C-domain 2]: Secreted.
Protein Families
Tumor necrosis factor family
Database Links
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