Neutralization Activity: This antibody has been reported to be useful for the neutralization of recombinant human IL-4. Neutralization may be measured with a proliferation assay using 2 ng/mL recombinant human IL-4 (Cat. No. 554605) to stimulate TF-1 cells at 1x10^5 cells/mL as indicator cells (i.e. preincubation of the antibody with recombinant human IL-4 for 60 minutes). 50% Neutralization (ND50) at 4 - 20 ng/mL >95% Neutralization at 20 - 100 ng/mL
Purification
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Sterility
0.2 μm filtered
Endotoxin Level
Endotoxin level is ≤ 0.01 EU/μg (≤ 0.001 ng/μg) of protein as determined by the LAL assay.
ELISA: Biotinylated MP4-25D2 antibody (Cat. No. 554483) has been reported to be useful as a detection antibody for sandwich ELISA measuring human IL-4 protein levels. Biotinylated MP4-25D2 antibody can be paired with the purified 8D4-8 antibody (Cat. No. 554515) as the capture antibody, with recombinant human IL-4 (Cat. No. 554605) as the standard. Investigators are encouraged to use the BD OptEIA™ Human IL-4 ELISA (Cat. No. 555194 or 550614) for measuring human IL-4 in serum or plasma samples.
Restrictions
For Research Use only
Format
Liquid
Concentration
1.0 mg/mL
Buffer
No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2μm sterile filtered.
Preservative
Azide free
Storage
4 °C
Storage Comment
Store undiluted at 4°C. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
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).
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).
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).
Kitamura, Tange, Terasawa, Chiba, Kuwaki, Miyagawa, Piao, Miyazono, Urabe, Takaku: "Establishment and characterization of a unique human cell line that proliferates dependently on GM-CSF, IL-3, or erythropoietin." in: Journal of cellular physiology, Vol. 140, Issue 2, pp. 323-34, (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.