16-7052-85

antibody from Invitrogen Antibodies

Targeting: IL5 EDF, IL-5, TRF

Provider product page for 16-7052-85

Immunohistochemistry

Blocking/Neutralizing

Other assay

Antibody data

Antibody Data
Antigen structure
References [5]
Comments [0]
Validations
Other assay [5]

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Product number: 16-7052-85 - Provider product page
Provider: Invitrogen Antibodies
Product name: IL-5 Monoclonal Antibody (TRFK5), Functional Grade, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The TRFK5 antibody reacts with mouse and human interleukin-5 (IL-5). The TRFK5 antibody is a neutralizing antibody. Mouse IL-5 is a disulfide-linked homodimer, containing two 113 amino acid peptides; these resolve as 32-34 kDa bands in SDS-PAGE. IL-5 is produced by T cells and has been known as eosinophil-differentiating factor (EDF), B cell growth factor II (BCGFII), and T cell-replacing factor (TRF). IL-5 induces eosinophil differentiation and promotes eosinophil survival and activation. In mice, IL-5 has been shown to stimulate B cell proliferation and antibody production. Applications Reported: The TRFK5 antibody has been reported for use in ELISA capture, ELISPOT capture, neutralization of IL-5 bioactivity, and IHC. Applications Tested: The Functional Grade Purified TRFK5 antibody has been tested by LAL assay to verify low endotoxin levels, by ELISA, and in bioassay for neutralization of mouse IL-5 bioactivity. The TRFK5 antibody at 0.06 µg/mL has been found to inhibit by 50% the biological effects of 1.0 ng/mL mouse IL-5 in an MC/9 cell proliferation assay. Detailed information and protocols about cytokine bioassays and in vitro cytokine neutralization using antibodies can be found in the BestProtocols® section. The TRFK5 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of mouse IL-5 in combination with the biotin TRFK4 (13-7051) antibody for detection and recombinant mouse IL-5 (14-8051) as the standard. A suitable range of concentrations of this antibody for ELISA capture is 1-4 µg/mL. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 1000 pg/mL - 8 pg/mL should be included in each ELISA plate.The TRFK5 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of human IL-5 in combination with the biotinylated JES1-5A10 (13-7059) antibody for detection and recombinant human IL-5 (14-80591) as the standard. A suitable range of concentrations of this antibody for ELISA capture is 1-4 µg/mL. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 1000 pg/mL - 8 pg/mL should be included in each ELISA plate. Storage and handling: Use in a sterile environment. Filtration: 0.2 µm post-manufacturing filtered. Purity: Greater than 90%, as determined by SDS-PAGE. Endotoxin Level: Less than 0.001 ng/µg antibody, as determined by LAL assay. Aggregation: Less than 10%, as determined by HPLC.
Reactivity: Human, Mouse
Host: Rat
Isotype: IgG
Antibody clone number: TRFK5
Vial size: 500 µg
Concentration: 1 mg/mL
Storage: 4° C

IL5 protein structure - 16-7052-85 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Naive mouse splenocytes (SJL/J strain) were stimulated with 2 µg/mL ConA in HL-1 media for 48 hours (right well) in a Mouse IL-5 ELISPOT assay plate. Left well is medium alone control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 Frequencies of CD4 + T-cells expressing Gata3 and Th2 cytokines in asthma and helminth infection are reduced in Bcl11b fl/fl dLck-iCre mice by an intrinsic defect. a Flow cytometry analysis of IL-4 (left column), IL-5 (middle column), and IL-13 (right column) against GATA3 in the CD4 + T-cell population of the lung parenchyma of HDM asthma-induced mice. b Frequencies of IL-4 + ( n = 4), IL-5 + ( n = 4), and IL-13 + ( n = 8) (top panel) and GATA3 + ( n = 10) (bottom panel) CD4 + T-cells from the lungs of the indicated groups. Data are derived from two (IL-4, IL-5, and IL-13 data) or five (GATA3 data) independent experiments. c Flow cytometry analysis of IL-4 (left column), IL-5 (middle column), and IL-13 (right column) against GATA3 in the CD4 + T-cell population from the mesenteric (m)LNs of H. polygyrus infected mice. d Frequencies of IL-4 + ( n = 4), IL-5 + ( n = 4), and IL-13 + ( n = 4) (top panel) and GATA3 + ( n = 8) (bottom panel) CD4 + T-cells from mLNs of the indicated groups. Data are derived from two (IL-4, IL-5, and IL-13) or four (GATA3) independent experiments. e Flow cytometry analysis of GATA3 in CD4 + T-cells (left column) and IL-4 (middle and right columns) in the GATA3 hi (middle column) and GATA3 lo (right column) CD4 + T-cells from the mLNs of H. polygyrus bakeri infected mice. f The ratio of GATA3 hi /GATA3 lo CD4 + T-cells (top) and frequencies of IL-4 secreting cells within the GATA3 hi and GATA3 lo CD4 + T-cells (bottom) from the mLNs of H. poly

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Influence of N. brasiliensis exposure on uncolonized FGT, increase in FGT eosinophils following Nb exposure (A) Female mice were hormone-synchronized 7 days prior to Nb infection. (B) At day 9 post-Nb infection (Nb 9dpi), levels of IL-33, IL-4, and IL-5 in FGT homogenates or lavages were assessed by ELISA or Luminex.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Nb-exacerbated HSV-2 pathology and FGT eosinophil infiltration is Il4ra independent WT and Il4ra -/- mice were infected with HSV-2 following Nb exposure as previously described. (A) HSV-2 progression was determined by daily pathology scoring. (B) Viral shedding was measured by plaque assay of day 6 vaginal washes. (C) Genital levels of IL-5 and IFN-gamma at day 2 post-HSV-2 infection, determined by Luminex and ELISA, respectively. Dotted line represents LLOQ of Luminex analysis. At 6 dpi, vaginal tissue was analyzed by H&E staining. (D) Representative sections (n = 3-4), displaying ulceration and inflammation of vaginal tissue. Images were taken at x50 magnification. HSV-2-ulcerated vaginal epithelium is indicated by black dotted lines and qualified as percentage (%) of ulcerated epithelium. (E) Representative Sirius-red-stained sections (n = 3-4) of virus-induced (Ei) epithelial ulcers and (Eii) stromal inflammation. Black arrows indicate eosinophil presence in the vaginal epithelial layer. Images were taken at x400 and x1,000 magnification. (F) Numbers (x10 3 ) of FGT eosinophils in WT and Il4ra -/- co-infected mice compared with HSV-2-only controls. Data are representative of two independent experiments with 3-6 mice per group (mean +- SEM). Statistical significance was calculated by two-way ANOVA with Bonferroni correction for multiple comparisons. * p