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Mouse TNF-alpha Recombinant



SKU: RKP06804 Tags: , , , , , , , , , ,


Optimized DNA sequence encoding Mouse Tumor Necrosis Factor-alpha extracellular domain was expressed in Escherichia Coli.
Molecular weight
Native Mouse Tumor Necrosis Factor-alpha is generated by the proteolytic removal of the signal peptide and propeptide, this molecule has a calculated mass of approximately 17kDa. Recombinant Mouse TNF is a monomer protein,consisting of 157 amino acids and migrates as an approximately17 kDa protein under reducing conditions.
>97%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determinedcytolysis of murine L929 cells in the presence of Actinomycin D is ≤.05 ng/ml, corresponding to a specific activity of ≥3 x units/mg.
Protein Sequence
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Recombinant mouse TNF alpha was lyophilized from.2 μm filtered PBS solution, pH7.0.
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Molecular function


Caspase 8 gene ablation provides neuroprotection in vitro in primary neuronal cultures and brain organotypic cultures.

  • 3-day-old PNC derived from embryos (E16.5) of CRE3 and Ncasp8

Transendothelial migration of mMSCs in response to chemokines.

  • MAECs were treated with or without TNFα and in the presence or absence of CXCL16, CXCL9, CCL20 and CCL25 (each at 10 ng/ml).


  • Tetramethylammonium hydroxide was from Shanghai Lingfeng Chemical Reagent Co Ltd, Shanghai, China.
  • Other materials were RPMI 1640 medium, penicillin, and streptomycin , fetal bovine serum (, , , the ), recombinant murine granulocyte-macrophage colony-stimulating factor, recombinant murine interleukin-4, CD11c, B7-1 (CD80), B7-2(CD86), MHC II, and chemokine receptor 7 (eBioscience, San Diego, CA), tumor necrosis factor-α, interleukin-1β, and interleukin-6 , prostaglandin E2 , a Prussian blue staining kit .
  • Annexin V and propidium iodide , and the Cell Counting Kit-8 .

Anti-inflammatory effects of diazoxide pre-treatment in microglial cell cultures stimulated with LPS and IFNγ.

  • Nitrite accumulation , and TNF-α and IL-6 release in control (unstimulated cells), DZX (unstimulated cells pretreated with 100 μM diazoxide), diazoxide and LPS/IFNγ + Diazoxide (10 μM to 100 μM) normalized for LPS/IFNγ untreated cells.

Flow cytometric analysis of endothelial VCAM-1 and E-selectin

  • Human umbilical vein endothelial cells were cultured in EGM-2 according to the manufacturer’s protocol.
  • HUVEC (passage 3–5) were plated in 24-well plates with 1×105 cells/400 µl per well in EGM-2 plus 20% FBS.
  • After a 5 hour re-attachment period, cells were pre-incubated for 16 hours with HDL isolated from pooled serum samples (n=6), after which TNF-α (0.5 ng/ml) was added to the culture medium for an additional 4 hours.
  • Cell-surface expression of VCAM-1 and E-selectin were then measured by flow cytometry, using a VCAM-1 monoclonal antibody followed by an anti-mouse FITC conjugated antibody , or an E-selectin PE conjugated antibody .
  • Cells were detached using 5 mM EDTA in PBS.
  • Cellular FITC and PE were then analyzed by flow cytometry (C6 Flow Cytometer, Accuri Cytometers ).
  • Controls included an isotype-matched control antibody and no primary antibody.
  • Native HDL3 (isolated by sequential ultracentrifugation of human plasma) at concentration of 0.5 mg/ml…

Epithelial cytokine expression and MSC migration.

  • B/ Supernatants from non-infected (white bars), 7.13- (light grey bars) or 26695- (dark grey bars) infected cells were assessed for TNFα and CCL2 expression by ELISA.

Enhanced circulating and intrathymic contents of TNF-α parallel the thymic atrophy in T. cruzi acutely-infected mice.

  • A) Representative picture showing two thymuses from healthy control mice (Co) and the progressive thymic atrophy after 14, 17 and 21 days of acute infection; B) TNF-α protein detection in thymus homogenates by western blot.

Less proinflammatory cytokine induction was evident in IL-1 KO mice after SCI.

  • TNFα level in the spinal cord of the wild-type (open squares and solid line) and IL-1 KO mice (closed square and dotted line).

PRP decreased the apoptotic cell death and caspase 3 activity induced by TNF-α and cycloheximide in ccdPAs.

  • The cells were treated with TNF-α and cycloheximide for 5, 30, 60, and 120 min in the medium with 2% FBS (closed bar) or 2% PRP (open bar).

Transendothelial Migration and Invadopodia (in Vitro Filipodia) Formation Assay of the T Lymphoma Cells

  • Trans-endothelial migration of T-cells was measured as described 6 cells) were seeded on the upper compartment on top of the endothelial monolayer in 100 µl.
  • After 16 hours of treatment, the tissue culture wells were removed and the lower compartment cells were collected to determine the number of transmigrated lymphocytes.
  • To assess the formation of leading edge filipodia, the T lymphoma cells were labelled with cell tracker green and added to confluent monolayers of tumour necrosis factor α (TNF-α) (cat 300-01A, , , ) activated bEnd.3 cells grown on glass cover slips that had been coated with 2 µg/ml fibronectin.
  • After, 16 hours of incubation, non adherent cells to the cover slips were removed by gentle washing the adherent cells were fixed with 4% paraformaldehyde and stained with DAPI and TRITC-phalloidin.
  • Z stack images were captured using Axiovision 4.1 Zeiss microscope and analyzed.
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