Mouse Tumor necrosis factor α,TNF-α ELISA KIT

Datasheet
  • 中文名称:
    小鼠肿瘤坏死因子α(TNF-α)ELISA KIT
  • 货号:
    CSB-E04741m
  • 规格:
    96T
  • 价格:
    3800
  • 其他:

产品详情

  • 产品描述:

    This Mouse Tumor necrosis factor α (TNF-α) ELISA KIT is used to quantitatively determine mouse TNF-α concentrations in serum, plasma, cell culture supernates, or tissue homogenates.TNF-α is a pleiotropic pro-inflammatory cytokine involved in multiple biological processes such as blood-brain barriers, brain-injury-related inflammatory, thrombogenic, and vascular alterations, immunological defense, and apoptosis. It exerts biological effects by binding to its main two receptors, TNFR1/p55 and TNFR2 /p75, on the cell surface.

    This kit employs the quantitative sandwich enzyme immunoassay technique to measure the levels of mouse TNF-α in the sample. Antibody specific for TNF-α has been pre-coated onto the microplate. Standards and samples are pipetted into the wells and any TNF-α present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated TNF-α antibody is added to the wells. After washing, avidin conjugated Horseradish Peroxidase (HRP) is added to the wells, forming an antibody-antigen-enzyme-labeled antibody complex. Following a wash to remove any unbound HRP-avidin, the TMB substrate solution is added to the wells and color develops in proportion to the amount of TNF-α bound in the initial step. The color changes from blue to yellow after the addition of stop solution into the wells. The absorbance (OD value) was determined by the microplate reader at the wavelength of 450nm, and the concentration of mouse TNF-α was calculated through the standard curve. Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, recovery, linearity, and lot-to-lot consistency. See the instructions for more information on validation.

  • 基因名:
    Tnf
  • 别名:
    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
  • 缩写:
    TNF
  • Uniprot No.:
  • 种属:
    Mus musculus (Mouse)
  • 样本类型:
    serum, plasma, cell culture supernates, tissue homogenates
  • 检测范围:
    15.6 pg/mL-1000 pg/mL
  • 灵敏度:
    3.9 pg/mL
  • 反应时间:
    1-5h
  • 样本体积:
    50-100ul
  • 检测波长:
    450 nm
  • 研究领域:
    Immunology
  • 测定原理:
    quantitative
  • 测定方法:
    Sandwich
  • 精密度:
    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.    
                 
  • 线性度:
    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  
  • 回收率:
    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  
                 
                 
  • 标准曲线:
    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 2.199 2.222 2.211 2.042  
    500 1.571 1.543 1.557 1.388  
    250 1.021 1.011 1.016 0.847  
    125 0.693 0.686 0.690 0.521  
    62.5 0.494 0.454 0.474 0.305  
    31.2 0.347 0.338 0.343 0.174  
    15.6 0.251 0.244 0.248 0.079  
    0 0.172 0.166 0.169    
  • 本试剂盒所含材料:
    • An assay plate --- The 96-well plate is composed of12 x 8 strip plates and can be taken apart.
    • Two bottles of lyophilized standard --- Reconstitute one bottle of the standard with sample diluent at dilution series and draw the standard curve.
    • Biotin-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.
    • Adhesive Strip (For 96 wells) 4 --- Seal the wells when incubation.
    • Instruction manual 1

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  • 本试剂盒不含材料:
    • 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

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  • 货期:
    3-5 working days

引用文献

问答及客户评论

 常见问题解答
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.

靶点详情

  • 功能:
    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.; FUNCTION
  • 基因功能参考文献:
    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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  • 亚细胞定位:
    Cell membrane, Single-pass type II membrane protein, SUBCELLULAR LOCATION: Tumor necrosis factor, membrane form: Membrane, Single-pass type II membrane protein, SUBCELLULAR LOCATION: Tumor necrosis factor, soluble form: Secreted, SUBCELLULAR LOCATION: C-domain 1: Secreted, SUBCELLULAR LOCATION: C-domain 2: Secreted
  • 蛋白家族:
    Tumor necrosis factor family
  • 数据库链接:

    KEGG: mmu:21926

    STRING: 10090.ENSMUSP00000025263

    UniGene: Mm.1293