Invitrogen

TNF alpha Monoclonal Antibody (MP6-XT22), Functional Grade, eBioscience™

Product Image Gallery

TNF alpha Antibody in Immunohistochemistry (IHC) — Figure 10 Coexpression of Fgr with p65, TNF-alpha, or ERK1/2 and coexpression of p65 with TNF-alpha in microglia of hemorrhagic thalami. Triple immunofluorescence staining showing coexpression of Iba1 with p-65 and Fgr, with TNF-alpha and Fgr, and with ERK1/2 and Fgr as well as of Fgr with p65 and TNF-alpha in the regions adjacent to the core of thalamic hemorrhagic injury on day 5 post-Coll IV microinjection. Representative images from 3 biological repeats ( n = 3 mice). Scale bar: 100 mum.

TNF alpha Antibody in Flow Cytometry (Flow) — Figure 6 Cytokines production after Fluvastatin treatment. (a) Splenic mononuclear cells from C57BL/6 mice treated with Fluvastatin or buffer for consecutive 7 days were harvested. Cells were cultured with PMA (50 ng/ mu l), ionomycin (1 mu g/ mu l) for 3-5 hours, and brefeldin A (5 mu g/ mu l) was added before flow cytometric analysis of TNF- alpha and IFN- gamma production in CD3 + T cells. One representative FACS data from 3 independent experiments was shown in the right panel, and the number in the corner represents the percentages of gated cells. (b) BALB/c mice were used as recipients following transplantation of Fluvastatin- or buffer-pretreated C57BL/6-derived bone marrow and splenic cells. Splenic cells from recipients were stimulated as described above, and then, the production of TNF- alpha and IFN- gamma in CD8 + and CD4 + T cells on days 7, 14, 21, and 28 were detected. Data represents the mean +- SD of results from 3 experiments ( n = 4 - 5 per time point). * P < 0.05, ** P < 0.01, n.s P > 0.05. (c) Representative FACS plots on day 21 were shown. The number in the corner means the cell percentages.

TNF alpha Antibody in Neutralization (Neu) — Fig. 5 PTP1B inhibition attenuates astrocytic TDP-43-induced toxicity and mitochondrial dysfunction in differentiated NSC-34 motor neuron-like cells. a - f NSC-34 cells were treated with 1 muM RA for 8 days and then were analysed. a CMFDA staining of undifferentiated or differentiated NSC-34 cells at 8 days. Scale bars, 100 mum. b Real-time PCR results of motor neuronal marker genes in undifferentiated or differentiated NSC-34 cells at the indicated time points. Data are presented as the mean +- SD. Gapdh was used for normalization. c Differentiated NSC-34 cells were stimulated with GFP ACM, TDP-43 ACM or TDP-43+ PTP1Bi ACM for 4 days. Cell viability was analysed by CCK-8 assay. *** p < 0.005 (one-way ANOVA). d Differentiated NSC-34 cells stimulated with TDP-43 ACM were treated with an IL-1beta antibody (50 ng/ml), an IL-6 antibody (50 ng/ml) and a TNF-alpha antibody (100 ng/ml) for 4 days and then were subjected to CCK-8 assays. TDP-43 ACM-induced toxicity was rescued by IL-1beta, IL-6 and TNF-alpha antibody treatment. Data are presented as the mean +- SD. * p < 0.05; ** p < 0.005 (one-way ANOVA). e, f Differentiated NSC-34 cells in XF24-well culture plates were stimulated with GFP ACM, TDP-43 ACM or TDP-43 + PTP1Bi ACM for 4 days. e Mitochondrial dysfunction analysis of ACM-treated cells was performed with a Seahorse XF analyser to detect the basal OCR, ATP production, maximum reserve and respiratory capacity. The OCR was normalized to total protein concentration. f Quantif

Product Details

Applications

Tested Dilution

Publications

Immunohistochemistry (IHC)

View 3 publications 3 publications

Flow Cytometry (Flow)

View 26 publications 26 publications

Neutralization (Neu)

Assay-Dependent

View 1 publication 1 publication

Functional Assay (FN)

Assay-Dependent

View 1 publication 1 publication

Product Specifications

Species Reactivity

Mouse

Published species

Mouse

Host/Isotype

Rat / IgG1, kappa

Recommended Isotype Control

Rat IgG1 kappa Isotype Control (eBRG1), Functional Grade, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

MP6-XT22

Conjugate

Functional Grade

Form

Liquid

Concentration

1 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

no preservative

Storage conditions

4° C

RRID

AB_494133

Product Specific Information

Description: The MP6-XT22 antibody reacts with mouse tumor necrosis factor-alpha (TNF-alpha), a 17 kDa cytokine produced by monocytes, macrophages, neutrophils, NK cells and CD4^+ T cells. TNF-alpha has cytolytic activity against a range of tumor cells and is important in immune regulation. TNF-alpha forms dimers and trimers and also exists as a 26 kDa membrane-bound form.

Applications Reported: The MP6-XT22 antibody has been reported for use in ELISA, neutralization, and immunohistochemistry.

Applications Tested: The Functional Grade Purified MP6-XT22 antibody has been tested by LAL assay to verify low endotoxin level and has been tested for neutralization of TNF alpha bioactivity.

The MP6-XT22 antibody at 1.25 µg/mL has been found to neutralize by 50% the biological effects of 1.0 ng/mL mouse TNF alpha in an assay of TNF alpha induced cytotoxicity of L929 cells. Detailed information and protocols about cytokine bioassays and in vitro cytokine neutralization using antibodies can be found in the BestProtocols® section.

It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Storage and handling: Use in a sterile environment.

Filtration: 0.2 µm post-manufacturing filtered.

Purity: Greater than 90%, as determined by SDS-PAGE.

Endotoxin Level: Less than 0.001 ng/µg antibody, as determined by LAL assay.

Aggregation: Less than 10%, as determined by HPLC.

Target Information

TNF alpha is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophage and bind to its receptors, TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. TNF alpha is involved in the regulation of immune cells, cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single 17 kDa band following SDS-PAGE under non-reducing conditions. Knockout studies in mice also suggested the neuroprotective function of TNF alpha, and has been observed to causes tumor necrosis when injected into tumor-bearing mice. Other functions of TNF-alpha include its role in the immune response to bacterial, viral, parasitic and certain fungal infections, as well as its role in the necrosis of specific tumors. TNF alpha causes cytolysis or cytostasis of certain transformed cells, being synergistic with interferon-gamma in its cytotoxicity. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: C-domain 1; C-domain 2; Cachectin; DADB-70P7.1; ICD1; ICD2; Intracellular domain 1; Intracellular domain 2; N-terminal fragment; NTF; TNF alpha; TNF superfamily; TNF α; TNF-a; TNF-alpha; TNFα; Tumor necrosis factor; Tumor necrosis factor ligand superfamily member 2; Tumor necrosis factor, membrane form; Tumor necrosis factor, soluble form; tumor necrosis factor-alpha

Gene Aliases: DIF; Tnf; TNF-a; TNF-alpha; Tnfa; TNFalpha; Tnfsf1a; TNFSF2

UniProt ID: (Mouse) P06804

Entrez Gene ID: (Mouse) 21926

Function(s)