TNFRSF1B
(Tumor Necrosis Factor Receptor Superfamily, Member 1B (TNFRSF1B))
Reactivity
Mouse
Host
Armenian Hamster
Clonality
Monoclonal
Conjugate
This TNFRSF1B antibody is un-conjugated
Application
Flow Cytometry (FACS), Immunoprecipitation (IP)
Brand
BD Pharmingen™
Characteristics
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results. 2. Please refer to us for technical protocols. 3. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. 4. 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
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
TNFRSF1B
Reactivity: Human
WB
Host: Rabbit
Monoclonal
unconjugated
Application Notes
Immunofluorescent Staining and Flow Cytometric Analysis: The purified form of TR75-89 (ABIN967654) can be used for the immunofluorescent staining (Less or equal than 1 µg antibody/10e6 cells) and flow cytometric analysis of normal mouse cells or cell lines to measure their expressed levels of p75 TNFR. An appropriate purified immunoglobulin isotype control is clone G235-2356. A three-layer staining protocol is recommended for maximizing the detection of p75 TNFR expressed by cells as detailed in the figure legend. TR75-89 is a nonblocking antibody that can be used for the unobstructed immunofluorescent staining and flow cytometric analysis of cells in systems where ligands (e.g., TNF) for p75 TNF receptors are present. Based on our testing results (data not shown), the presence of exogenous recombinant mouse TNF at levels Less or equal than 1 µg per 10e6 cells was insufficient to inhibit the binding of TR75-89 to L929 cells that express p75 TNFR (at 0.25 µg antibody/10e6 cells). Please note also that as a consequence of in vivo or in vitro activation, cell surface p75 TNFR can either be shed by cells or transiently expressed at higher levels. As a result, cellular activation can affect the cell's overall expressed level of surface p75 TNFR. IP: The TR75-89 antibody has been reported to be useful for the immunoprecipitation of p75 TNFR from lysates of mouse Meth A fibrosarcoma cells. Please note that this application is not routinely tested.
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C
Storage Comment
Store undiluted at 4° C.
Zola: "Detection of cytokine receptors by flow cytometry." in: Current protocols in immunology / edited by John E. Coligan ... [et al.], Vol. Chapter 6, pp. Unit 6.21, (2008) (PubMed).
Pinckard, Sheehan, Arthur, Schreiber: "Constitutive shedding of both p55 and p75 murine TNF receptors in vivo." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 158, Issue 8, pp. 3869-73, (1997) (PubMed).
Pinckard, Sheehan, Schreiber: "Ligand-induced formation of p55 and p75 tumor necrosis factor receptor heterocomplexes on intact cells." in: The Journal of biological chemistry, Vol. 272, Issue 16, pp. 10784-9, (1997) (PubMed).
Sheehan, Pinckard, Arthur, Dehner, Goeddel, Schreiber: "Monoclonal antibodies specific for murine p55 and p75 tumor necrosis factor receptors: identification of a novel in vivo role for p75." in: The Journal of experimental medicine, Vol. 181, Issue 2, pp. 607-17, (1995) (PubMed).
Target
TNFRSF1B
(Tumor Necrosis Factor Receptor Superfamily, Member 1B (TNFRSF1B))
The TR75-89 antibody reacts with the extracellular region of CD120b, the 75 kDa receptor for the mouse cytokines, tumor necrosis factor (TNF, aka TNF-alpha) and lymphotoxin-alpha (LT-alpha, aka lymphotoxin, TNF-beta). This receptor, referred to as the p75 or Type II Tumor Necrosis Factor Receptor (TNFRII), or TNFRSF1B, is expressed by a variety of cell lines and normal cell types including T cells, monocytes, macrophages, and neutrophils. Resting B cells express low or undetectable levels of TNFRII whereas mature erythrocytes are uniformly negative for TNFRII expression. In addition, the TR75-89 antibody can bind to a soluble, truncated form of the mouse Type II TNFR that is shed by cells in response to certain stimuli, e.g., cells treated with LPS or TNF. The in vivo administration of nonblocking, nonagonistic TR75-89 antibody reportedly results in the linear accumulation of shed, soluble forms of p75 TNFR in the circulation. The TR75-89 antibody does not recognize the 55 kDa (p55) Type I TNFR (aka, CD120a). TR75-89 does not block the binding and does not neutralize the bioactivity of the TNF ligand on L929 target cell populations that express p75 (and p55) TNFR. The immunogen used to generate the TR75-89 hybridoma was a purified, soluble extracellular domain of the mouse Type II TNFR. This antibody is routinely tested by flow cytometric analysis. Synonyms: TNF Receptor, Type II/p75