TNF alpha Protein

Catalog No. ABIN1305135

This Recombinant TNF alpha protein is expressed in Escherichia coli (E. coli) and has been mentioned in 3+ publications.

Quick Overview for TNF alpha Protein (ABIN1305135)

Target: TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))

Protein Type: Recombinant

Biological Activity: Active

Origin: Mouse

Source: Escherichia coli (E. coli)

Application: ELISA, Functional Studies (Func), Intracellular Flow Cytometry (ICFC), Blocking Reagent (BR)

Purity: ≥ 95 %

Product Details

Brand: BD Pharmingen™

Specificity: Biological Activity: Activity Test: MTT/Actinomycin D Cytolysis assay using L929 indicator cells at 2 x 10^5 cells/ml
Specific Activity: 0.2 - 5.0 x 10^9 Units /mg
ED50: 2 - 50 pg/ml
Observed dose response relationship: 80 fold A Unit is defined as the amount of material required to stimulate a half-maximal response at cytokine saturation. Since applications vary, each investigator should titrate the reagent to obtain optimal results.

Characteristics: Recombinant mouse TNF is supplied as a frozen aqueous buffered solution containing biotechnology grade, low endotoxin bovine serum albumin, with no preservatives. Recombinant mouse TNF was purified and found to be > 95% pure by SDS-PAGE, and an absorption assay based on the Beers-Lambert Law.

Purification: Recombinant mouse TNF is supplied as a frozen liquid comprised of 0.22 μm sterile-filtered aqueous buffered solution containing bovine serum albumin, with no preservatives. Recombinant mouse TNF is ≥ 95 % pure as determined by SDS-PAGE, and an absorption assay based on the Beers-Lambert Law.

Sterility: 0.22 μm filtered

Endotoxin Level: The endotoxin level is ≤ 0.1 ng per µg of TNF, as measured in a chromogenic LAL assay.

Application Details

Application Notes: ELISA Standard: Recombinant mouse TNF is useful as a quantitative standard for measuring mouse TNF protein levels in an TNF specific sandwich ELISA with the purified G281-2626 antibody (Cat. No. 551225) as a capture antibody and the biotinylated MP6-XT3 (Cat. No. 554415) as the detection antibody. To obtain linear standard curves, doubling dilutions of this mouse TNF standard from ~2,000 to 15 pg/ml should be included in each ELISA plate.
Note: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assaying serum or plasma samples. For measuring mouse TNF in serum or plasma the BD OptEIA™ Mouse TNF ELISA Sets (Cat. No. 555268 or 558534) and BD OptEIA™ Mouse TNF ELISA Kit (Cat. No. 559732 or 560478) are specially formulated and recommended.
Ligand Blocking Control for Immunfuorescent Staining of Cytokines: Recombinant mouse TNF can be used as a blocking control to demonstrate specificity of TNF staining by the directly conjugated-MP6-XT22 antibody. To perform this control, the fluorochrome-conjugated antibody can be preincubated with 0.05 - 0.5 µg of recombinant mouse TNF for 20 minutes at 4°C, prior to staining. The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.

Comment:

BD Pharmingen™ Recombinant Mouse TNF - Reactivity Ms

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 200 μg/mL

Buffer: Frozen aqueous buffered solution containing BSA.

Preservative: Azide free

Handling Advice: This preparation contains no preservatives, thus it should be handled under aseptic conditions. Rapidly thaw and quick-spin product prior to use. Upon initial thawing, the product should be aliquoted into polypropylene microtubes and frozen at -80°C for future use. Alternatively, the product can be diluted in sterile neutral buffer containing not less than 0.5 - 10 mg/ml carrier protein such as human or bovine serum albumin, aliquoted and stored at -80°C. For in vitro biological assay use, we recommend carrier protein concentrations of ≥ 1 mg/ml. For use as an ELISA standard, we recommend carrier protein concentrations of 5 - 10 mg/ml. The product should not be diluted to less than 50 µg/ml for long term storage. Failure to add carrier protein or store at indicated temperatures may result in a loss of activity.
Note: Carrier proteins should be pre-screened for possible effects in an appropriate experimental system. Carrier proteins may effect exper imental results due to toxicity, high endotoxin levels or possible blocking activity.

Storage: -80 °C

Storage Comment: Store product at -80°C prior to use or for long term storage of stock solutions. Rapidly thaw and quick-spin product prior to use. Avoid multiple freeze-thaws of product. This preparation contains no preservatives, thus it should be handled under aseptic conditions.

References

Prussin, Metcalfe: "Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies." in: Journal of immunological methods, Vol. 188, Issue 1, pp. 117-28, (1996) (PubMed).

Jäättelä: "Biologic activities and mechanisms of action of tumor necrosis factor-alpha/cachectin." in: Laboratory investigation; a journal of technical methods and pathology, Vol. 64, Issue 6, pp. 724-42, (1991) (PubMed).

Hogan, Vogel: "Production of tumor necrosis factor by rIFN-gamma-primed C3H/HeJ (Lpsd) macrophages requires the presence of lipid A-associated proteins." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 141, Issue 12, pp. 4196-202, (1989) (PubMed).

Target Details

Target: TNF alpha (Tumor Necrosis Factor alpha (TNF alpha))

Alternative Name: TNF

Background: Tumor Necrosis Factor (TNF, aka TNF-α) is a potent multifunctional cytokine which can exert regulatory and cytotoxic effects on a wide range of normal lymphoid and non-lymphoid cells and tumor cells. Mouse TNF is a differentially glycosylated protein containing 156 amino acid residues.

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