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Il17a Antibody (OASA08429)

Description of Target:
RABBIT ANTI MOUSE INTERLEUKIN-17:Biotin
Gene Symbol:
Il17a
NCBI Gene Id:
16171
Alias Symbols:
Il17; Ctla8; Ctla-8; Il17a
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express Il17a.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express Il17a.
Swissprot Id:
Q62386
Protein Accession #:
NP_034682.1
Host:
Rabbit
Protein Name:
Interleukin-17A
Protein Size (# AA):
158
Specificity:
IL-17
Application:
ELISA, WB
Conjugation:
Biotin
Immunogen:
The immunogen for anti-Il17a antibody: recombinant mouse IL-17
Product Format:
Purified IgG conjugated to Biotin - lyophilised
Isotype:
Polyclonal IgG
Predicted Homology Based on Immunogen Sequence:
Mouse
Species Reactivity:
Mouse
Datasheets / Downloads:
Printable datasheet for
anti-Il17a antibody
- OASA08429
Applications Info:
ELISA : This antibody may be used in an indirect ELISA or as a detection reagent in a sandwich ELISA with OASA08428 as the capture antibody.
:::
Preservative Stabilisers: None present
Antiserum Preparation: Antiserum to mouse IL-17 were raised by repeated immunisation of rabbits with highly purified antigen. Purified IgG was prepared by affinity chromatography.
:::
Approx Protein Conc: IgG concentration 0.1mg/ml after reconstitution
Buffer Solutions: Phosphate buffered saline pH7.2
Reconstitution and Storage:
Reconstitution: Reconstitute with 0.5ml sterile PBS containing 0.1% bovine serum albumin
Care should be taken during reconstitution as the protein may appear as a film at the bottom of the vial. Aviva Systems Biology recommend that the vial is gently

Product Protocol: Il17a antibody used to evaluate endogenous il-17 as a mediator of neutrophil recruitment caused by endotoxin exposure in mouse airways (OASA08429)

Specificity: IL-17
Product Page: Il17a antibody (OASA08429)
Antibody Pair Available: OASA08428 Capture OASA08429 Detection
Experiment Type: Detection of IL-17 protein with ELISPOT

Protocol:
Detection of soluble mIL-17 protein released from a coculture of positively selected lung CD3+ cells plus airway macrophages was conducted using ELISPOT assay. Briefly, cell culture plates (M200; ImmunoSpot, Cleveland, OH) were coated overnight at 4°C with a monoclonal anti-mouse capture Ab (clone 50101.111; 5.0 μg/ml; R&D Systems) dissolved in PBS. The plates were then blocked with blocking solution (RPMI 1640 plus 10% FCS plus 100 U/ml penicillin, 100 U/ml streptomycin, all from Sigma-Aldrich).

CD3+ lymphocytes from lung tissue were enriched over a magnetic field. The cells were seeded (0.5 × 106 cells/well) in the plates with and without airway macrophages, respectively, in complete medium (RPMI 1640 plus 10% FCS plus 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml sodium-pyruvate, and 4 mM l-glutamine). One-half of the cultures were stimulated with CI (1 μg/ml) and PMA (2 ng/ml). After 20 h of incubation in 5% CO2 at 37°C, the plates were washed (PBS plus 0.05% Tween 20), and the biotinylated anti-mIL-17 (clone AUS01; 2 μg/ml; R&D Systems) was used to detect the captured extracellular IL-17 protein. Spots were visualized using AV-HPR enzyme and 3-amino-9-ethylcarbazole substrate. The plates were washed in water to stop the reaction. Spots were enumerated using a dissecting microscope at a magnification of ×40 (CETI, Antwerpen, Belgium).

Summary:
1. As analyzed with ELISPOT assay (cells pooled from seven mice, used in three to five parallel experiments), positively selected CD3+ lung lymphocytes cocultured with airway macrophages displayed a substantial increase in released soluble mIL-17 protein in response to CI plus PMA (CI plus PMA: 19.0 (2.8) spots compared with 6.0 (1.6) spots for vehicle, p < 0.01). The monoculture of positively selected CD3+ lung lymphocytes did not display any substantial release of soluble mIL-17 protein (CI plus PMA: 7.2 (1.7) spots compared with 3.7 (1.2) spots for vehicle, p > 0.05). 2. Using the same ELISPOT assay, preliminary experiments indicated that airway macrophages in monoculture were unable to release soluble mIL-17 protein after stimulation with CI plus PMA (data not shown).

References:
1: Miyamoto M, Prause O, Sjöstrand M, Laan M, Lötvall J, Lindén A. Endogenous IL-17 as a mediator of neutrophil recruitment caused by endotoxin exposure in mouse airways. J Immunol. 2003 May 1;170(9):4665-72. PubMed PMID: 12707345.

 
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