Neutralization Activity: This antibody has been reported to be useful for the neutralization of recombinant human IL-5. Neutralization activity may be measured with a proliferation assay using 1.5 ng/mL recombinant human IL-5 (Cat. No. 554606) to stimulate TF-1 cells at 1x10^5 cells/mL as indicator cells (i.e. preincubation of the antibody with recombinant human IL-5 for 60 minutes). 50% Neutralization (ND50) at 5 - 50 ng/mL >95% Neutralization at 50 - 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.
IL5
Reactivity: Human
ELISA, D
Host: Rat
Monoclonal
JES1-5A10
Biotin
Application Notes
ELISA: The purified format of the TRFK5 antibody (Cat. No.554393) has been reported to be useful as a capture antibody for sandwich ELISA for measuring mouse or human IL-5. For a mouse IL-5 ELISA, purified TRFK5 antibody can be paired with the biotinylated TRFK4 antibody (Cat. No. 554397) as the detection antibody, with recombinant mouse IL-5 (Cat. No. 554581) as the standard. Alternatively, investigators may be interested in the BD OptEIA™ Mouse IL-5 ELISA Set (Cat. No. 555236). For a human IL-5 ELISA, purified TRFK5 antibody can be paired with the biotinylated JES1-5A10 antibody (Cat. No. 554491) as the detecting antibody, with recombinant human IL-5 (Cat. No. 554606) as the standard. Alternatively, investigators may be interested in the BD OptEIA™ Human IL-5 ELISA Set (Cat. No 555202).
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.
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).
Mo, Sarawar, Doherty: "Induction of cytokines in mice with parainfluenza pneumonia." in: Journal of virology, Vol. 69, Issue 2, pp. 1288-91, (1995) (PubMed).
Litton, Sander, Murphy, OGarra, Abrams: "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).
Sarawar, Sangster, Coffman, Doherty: "Administration of anti-IFN-gamma antibody to beta 2-microglobulin-deficient mice delays influenza virus clearance but does not switch the response to a T helper cell 2 phenotype." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 153, Issue 3, pp. 1246-53, (1994) (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).
Kitamura, Takaku, Miyajima: "IL-1 up-regulates the expression of cytokine receptors on a factor-dependent human hemopoietic cell line, TF-1." in: International immunology, Vol. 3, Issue 6, pp. 571-7, (1991) (PubMed).
Sander, Andersson, Andersson: "Assessment of cytokines by immunofluorescence and the paraformaldehyde-saponin procedure." in: Immunological reviews, Vol. 119, pp. 65-93, (1991) (PubMed).
Swain, Weinberg, English, Huston: "IL-4 directs the development of Th2-like helper effectors." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 145, Issue 11, pp. 3796-806, (1991) (PubMed).
Schumacher, OGarra, Shrader, van Kimmenade, Bond, Mosmann, Coffman: "The characterization of four monoclonal antibodies specific for mouse IL-5 and development of mouse and human IL-5 enzyme-linked immunosorbent." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 141, Issue 5, pp. 1576-81, (1988) (PubMed).
The TRFK5 antibody reacts with mouse interleukin-5 (IL-5) and cross-reacts with human IL-5. The TRFK5 antibody has been reported to cross react with IL-5 from rhesus monkey. This is a neutralizing antibody.