Tumor Necrosis Factor (TNF) antibody

Details for Product No. ABIN967462
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Antigen
Synonyms TNF, tnf, dif, tnfa, xtnf, TNF-a, tnfsf2, tnf-alpha, TNF-alpha, DIF, TNFA, TNFSF2, TNFalpha, Tnfa, Tnfsf1a, RATTNF, TNFa, cTNF, TNF-ALPHA, Cachectin, Tnf-alpha, tnfa-like
Reactivity
Mouse (Murine)
(495), (209), (98), (69), (64), (35), (23), (22), (10), (10), (8), (5), (5), (4), (4), (4), (2), (2), (1), (1)
Host
Rat
(330), (304), (97), (25), (13), (12), (9), (4), (4)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(86), (49), (24), (11), (8), (6), (6), (6), (4), (4), (4), (4), (4), (4), (3), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Flow Cytometry (FACS), ELISA (Capture), Western Blotting (WB)
(458), (400), (214), (111), (94), (87), (69), (61), (58), (52), (39), (31), (30), (20), (19), (6), (5), (5), (4), (3), (3), (3), (3), (3), (2), (2), (1), (1), (1)
Pubmed 5 references available
Quantity 0.1 mg
Options
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Catalog No. ABIN967462
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Immunogen Recombinant mouse TNF
Clone MP6-XT22
Isotype IgG1
Characteristics 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Alternative Name TNF
Background The MP6-XT22 antibody reacts with mouse tumor necrosis factor (TNF, also known as TNF-alpha). The immunogen used to generate this hybridoma was recombinant mouse TNF. This antibody is routinely tested by flow cytometric analysis.
Application Notes Blocking Control for Intracellular Staining:
The purified MP6-XT22 antibody (ABIN967462) can be used as a blocking control to demonstrate specificity of TNF protein staining by directly conjugated -MP6-XT22 antibody. To perform this control, the fixed/permeabilized cells (~ 1 million) can be incubated with 1 -10 µg of unlabeled MP6-XT22 antibody (ABIN967462) for 20 minutes at 4°C, prior to staining with directly conjugated antibody (e.g., 0.1 -0.5 µg mAb/1 million cells). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.

ELISA Capture:
The purified MP6-XT22 antibody (ABIN967462) is useful as a capture antibody for a sandwich ELISA for measuring mouse TNF protein levels. Purified MP6-XT22 antibody can be paired with the biotinylated polyclonal rabbit anti-mouse/rat TNF antibody as the detecting antibody, with recombinant mouse TNF as the standard. This pair measures total TNF, free TNF as well as TNF bound by soluble receptors.
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.

WB:
The MP6-XT22 antibody has been reported to be useful for Western blotting. Please note that this application is not routinely tested.
Restrictions For Research Use only
Format Liquid
Concentration 0.5 mg/ml
Buffer Aqueous buffered solution.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage 4 °C
Supplier Images
anti-Tumor Necrosis Factor (TNF) antibody Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. After 6 hour stimulation with hamster anti-mouse CD3 (2 µg/ml final concentration, Clone 145-2C11) and hamster anti-mouse CD28 (2 µg/ml final concentration, Clone 37.51) antibodies in the presence of GolgiStop™ (2 µM final concentration), the splenocytes were stained with FcBlock™ (1 µg/1 million cells), then 0.06 µg of PE-conjugated rat anti-mouse CD4 (PE-RM4-5). The cells were then fixed, permeabilized, and subsequently stained with 0.06 µg of FITC-conjugated rat anti-mouse TNF antibody (FITC-MP6-XT22) by using Pharmingen's staining protocol (first panel). To demonstrate specificity of staining, the binding of the FITC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF (0.25 µg, middle panel), and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody (2 µg, ABIN967462, second panel) prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking (middle panel) and antibody blocking (second panel) specificity controls.
anti-Tumor Necrosis Factor (TNF) antibody (2) The binding of the FITC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF (0.25 µg, middle panel)
anti-Tumor Necrosis Factor (TNF) antibody (3) Preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody (2 µg, ABIN967462, second panel)
Product cited in: Abrams: "Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies." in: Current protocols in immunology / edited by John E. Coligan ... [et al.], Vol. Chapter 6, pp. Unit 6.20, 2008 (PubMed).

General Abrams, Roncarolo, Yssel et al.: "Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples." in: Immunological reviews, Vol. 127, Issue 9-10, pp. 5-24, 1992 (PubMed).

Masaki, Kano, Merrick et al.: "Studies on Paul-Bunnell (P-B) antigen-antibody system. II. P-B antigens in extracts of lymphoma-leukemia spleens and pathologic sera." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 127, Issue 1, pp. 5-8, 1981 (PubMed).

Litton, Sander, Murphy et al.: "Early expression of cytokines in lymph nodes after treatment in vivo with Staphylococcus enterotoxin B." in: Journal of immunological methods, Vol. 175, Issue 1, pp. 47-58, 1994 (PubMed).

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).

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