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TNF alpha ELISA Kit

TNF alpha Reactivity: Mouse AA 80-235 Colorimetric Sandwich ELISA 15.6-1000 pg/mL Cell Culture Supernatant, Plasma (EDTA), Plasma (heparin), Serum
Catalog No. ABIN411363
  • Target See all TNF alpha ELISA Kits
    TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))
    Binding Specificity
    AA 80-235
    Reactivity
    • 14
    • 12
    • 8
    • 5
    • 4
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Mouse
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    15.6-1000 pg/mL
    Minimum Detection Limit
    15.6 pg/mL
    Application
    ELISA
    Purpose
    Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse TNF alpha
    Brand
    PicoKine™
    Sample Type
    Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)
    Analytical Method
    Quantitative
    Specificity
    Expression system for standard: E.coli,L80-L235
    Cross-Reactivity (Details)
    There is no detectable cross-reactivity with other relevant proteins.
    Sensitivity
    <1pg/mL
    Material not included
    Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl
    Immunogen
    Expression system for standard: E.coli
    Immunogen sequence: L80-L235
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  • Application Notes
    Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.
    Plate
    Pre-coated
    Protocol
    mouse TNF alpha ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for TNF alpha has been precoated onto 96-well plates. Standards (E.coli,L80-L235) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for TNF alpha is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse TNF alpha amount of sample captured in plate.
    Assay Procedure

    Aliquot 0.1 mL per well of the 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL, 15.6pg/mL mouse TNF alpha standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin , EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse TNF alpha standard solution and each sample be measured in duplicate.

    Assay Precision
    • Sample 1: n=16, Mean(pg/ml): 126, Standard deviation: 5.42, CV(%): 4.3
    • Sample 2: n=16, Mean(pg/ml): 228, Standard deviation: 14.82, CV(%): 6.5
    • Sample 3: n=16, Mean(pg/ml): 613, Standard deviation: 35, CV(%): 5.7,
    • Sample 1: n=24, Mean(pg/ml): 135, Standard deviation: 7.02, CV(%): 5.2
    • Sample 2: n=24, Mean(pg/ml): 241, Standard deviation: 16.87, CV(%): 7
    • Sample 3: n=24, Mean(pg/ml): 747, Standard deviation: 50.05, CV(%): 6.7
    Restrictions
    For Research Use only
  • Handling Advice
    Avoid multiple freeze-thaw cycles.
    Storage
    -20 °C,4 °C
    Storage Comment
    Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles
    Expiry Date
    12 months
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  • Target See all TNF alpha ELISA Kits
    TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))
    Alternative Name
    TNF (TNF alpha Products)
    Synonyms
    DIF ELISA Kit, TNF-alpha ELISA Kit, TNFA ELISA Kit, TNFSF2 ELISA Kit, RATTNF ELISA Kit, Tnfa ELISA Kit, tnf ELISA Kit, TNF-a ELISA Kit, TNFalpha ELISA Kit, Tnfsf1a ELISA Kit, TNFa ELISA Kit, cTNF ELISA Kit, Tnf-alpha ELISA Kit, tnfa-like ELISA Kit, TNF-ALPHA ELISA Kit, dif ELISA Kit, tnfa ELISA Kit, xtnf ELISA Kit, tnfsf2 ELISA Kit, tnf-alpha ELISA Kit, Cachectin ELISA Kit, tumor necrosis factor ELISA Kit, tumor necrosis factor b (TNF superfamily, member 2) ELISA Kit, tumor necrosis factor alpha ELISA Kit, tumor necrosis factor a (TNF superfamily, member 2) ELISA Kit, TNF ELISA Kit, Tnf ELISA Kit, tnf ELISA Kit, tnfb ELISA Kit, tnf-alpha ELISA Kit, LOC103694380 ELISA Kit, tnfa ELISA Kit
    Background

    Protein Function: 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.

    Background: Tumor necrosis factor-alpha(TNF-alpha, or TNF) is secreted by macrophages in response to inflammation, infection and cancer. Human Tumor Necrosis Factor(TNF) and Lymphotoxin(TNF-beta) are cytotoxic proteins which have similar biological activities and share 30 % amino acid homology. TNF-alpha is produced by monocytes, which can stimulate endothelial cells to produce the multilineage growth factor granulocyte-macrophage colony-stimulating factor and extend the role of this immunoregulatory protein to the regulation of hematopoiesis in vitro. TNF is a soluble protein that causes damage to tumor cells but has no effect on normal cells. Human TNF has been purified to apparent homogeneity as a 17.3-kilodalton protein from HL-60 leukemia cells and has showed cytotoxic and cytostatic activities against various human tumor cell lines. The human TNF cDNA is 1585 base pairs in length and encodes a protein of 233 amino acids. The mature protein begins at residue 77, leaving a long leader sequence of 76 amino acids. TNF-alpha has been mapped to human chromosome 6.

    Synonyms: Tumor necrosis factor,Cachectin,TNF-alpha,Tumor necrosis factor ligand superfamily member 2,TNF-a,Tumor necrosis factor, membrane form,N-terminal fragment,NTF,Intracellular domain 1,ICD1,Intracellular domain 2,ICD2,C-domain 1,C-domain 2,Tumor necrosis factor, soluble form,Tnf,Tnfa, Tnfsf2,

    Full Gene Name: Tumor necrosis factor

    Cellular Localisation: Cell membrane, Single-pass type II membrane protein.
    Gene ID
    21926
    UniProt
    P06804
    Pathways
    NF-kappaB Signaling, Apoptosis, Caspase Cascade in Apoptosis, TLR Signaling, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus, Inflammasome
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