Western Blotting (WB), Blocking Antibody (Inhibition)
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. 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.
1. Blocking Control for Intracellular Staining: The unlabeled MP4-25D2 antibody can be used as a blocking control to demonstrate specificity of IL-4 staining by conjugated MP4-25D2 antibody. To perform this control, the fixed/permeabilized cells (~ 1 million) can be incubated with 1-10 µg of unlabeled MP4-25D2 antibody (ABIN967463) for 20 minutes at 4°C, prior to staining with conjugated MP4-25D2 antibody. The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.
2. Neutralization: The NA/LE™ MP4-25D2 antibody is useful for neutralization of human IL-4 bioactivity. A suitable NA/LE ™ rat IgG1 isotype control to match the MP4-25D2 antibody is the R3-34 antibody.
3. WB: The purified MP4-25D2 antibody has been reported to be useful for Western blotting. 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.
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).
Jung, Schauer, Rieger, Wagner, Einsle, Neumann, Heusser: "Interleukin-4 and interleukin-5 are rarely co-expressed by human T cells." in: European journal of immunology, Vol. 25, Issue 8, pp. 2413-6, (1995) (PubMed).
Ramanathan, Ingram, Sullivan, Greenberg, Reim, Trotta, Le: "Immunochemical mapping of domains in human interleukin 4 recognized by neutralizing monoclonal antibodies." in: Biochemistry, Vol. 32, Issue 14, pp. 3549-56, (1993) (PubMed).
Abrams, Roncarolo, Yssel, Andersson, Gleich, Silver: "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).
Chrétien, Van Kimmenade, Pearce, Banchereau, Abrams: "Development of polyclonal and monoclonal antibodies for immunoassay and neutralization of human interleukin-4." in: Journal of immunological methods, Vol. 117, Issue 1, pp. 67-81, (1989) (PubMed).
The MP4-25D2 antibody reacts with human interleukin-4 (IL-4). The immunogen used to generate the MP4-25D2 hybridoma was purified recombinant human IL-4. This is a neutralizing antibody. The MP4-25D2 antibody has been reported to cross react with IL-4 from rhesus monkeys. The use of the MP4-25D2 antibody for epitope mapping of human IL-4 has been described. This antibody is routinely tested as a blocking control for intracellular staining.