anti-IL-17A antibody product blog
Tags: Antibody; IL-17A; Monoclonal Antibody; anti-IL-17A antibody;
The IL-17A il17a (Catalog #MBS8505777) is an Antibody and is intended for research purposes only. The product is available for immediate purchase. The Mouse anti-Human IL-17A Monoclonal Antibody, Purified reacts with Human and may cross-react with other species as described in the data sheet. MyBioSource\'s IL-17A can be used in a range of immunoassay formats including, but not limited to, ICFC. Researchers should empirically determine the suitability of the IL-17A il17a for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process.The IL-17A il17a product has the following accession number(s) (GI #4504651) (NCBI Accession #NP_002181.1) (Uniprot Accession #Q16552). Researchers may be interested in using Bioinformatics databases such as those available at The National Center for Biotechnology Information (NCBI) website for more information about accession numbers and the proteins they represent. Even researchers unfamiliar with bioinformatics databases will find the NCBI databases to be quite user friendly and useful.
To buy or view more detailed product information and pricing, please click on the technical datasheet page below:
IL-17A is the founding member of the IL-17 family, a group of six structurally related pro-inflammatory cytokines. IL-17A, secreted by activated CD4+ Th17 cell subpopulation, elicits multiple biological activities on a variety of cells including: the induction of IL-6, IL-8, G-CSF, and PGE2 production in epithelial, endothelial or fibroblasts; the enhancement of surface expression of ICAM-1 in fibroblasts; activation of NF-?B and costimulation of T cell proliferation. Recent studies demonstrated that, in mice, activated IL-17-secreting CD4+ helper T cells (Th17 cells) mediate an autoimmune arthritis that clinically and immunologically resembles rheumatoid arthritis (RA). Human IL-17A shows 63%, 63%, and 72% amino acid sequence identity to rat IL-17A, mouse IL-17A, and a protein encoded by the ORF13 gene of herpesvirus Saimiri (HVS), respectively.
Experimental Methods: The following is a general protocol, optimization is required for best results of your experiment.
1. Use Human Mononuclear Cells Separation Medium to harvest PBMCs from heparinized whole blood.
[Human Mononuclear Cells Separation Medium: Cat.: E16-25610]
2. Add appropriate amount of cell stimulation cocktail and protein transport inhibitor cocktail, and incubate for 4-6 hours in 5% CO2 at 37�C.( the stimulation time for different cytokines is different.)
[Cell Stimulation Cocktail: Cat.: E16-FXP048]
[Protein Transport Inhibitor Cocktail: Cat.: E16-FXP050]
[Cell Stimulation Cocktail plus protein transport inhibitors: Cat.: E16-FXP049]
3. Sample tube is set to 1000 rpm centrifugation for 5 minutes, discard the supernatant; 4. Add 2 ml PBS wash buffer to resuspend the cells, then 1000 rpm centrifugation for 5 minutes, discard the supernatant; Perform cell surface staining
5. Add 0.5 ml PBS wash buffer to resusend the cells.
6. Add appropriate amount of cell surface antibodies to the bottom of flow tube mixing with the PBMCs, incubate for 20 minutes at room temperature away from light;
7. Add 2 ml PBS wash buffer to resuspend the cells, then1000 rpm centrifugation for 5 minutes, discard the supernatant; Perform Cytokines antibody staining
8. Resuspend cells in 0.5 ml/tube Fixation Buffer and incubate 10min at RT(recommended: Fixation Buffer,Cat.: E16-FXP008);
9. Sample tube is set to 1000 rpm centrifugation for 5 minutes, discard the supernatant;
10. Add 2 ml PBS wash buffer to resuspend the cells, then 1000 rpm centrifugation for 5 minutes, discard the supernatant;
11. Resuspend fixed cells in 1.5 ml/tube Permeabilization Buffer and incubate 10min at RT(recommended: Permeabilization Buffer,Cat.: E16-FXP009);
12. Sample tube is set to 1000 rpm centrifugation for 5 minutes, discard the supernatant;
13. Resuspend fixed/permeabilized cells in the approximate 100 μl Permeabilization Buffer and add appropriate amount of purified anti-human IL-17A antibody to the bottom of flow tube, incubate for 20 minutes at room temperature;
14. Add 2 ml PBS wash buffer to resuspend the cells, then 1000 rpm centrifugation for 5 minutes, discard the supernatant;
15. Add appropriate amount of fluorescent-labeled anti-mouse IgGs and incubate for 20 minutes away from light at room temperature.
16. Sample tube is set to 1000 rpm centrifugation for 5 minutes, discard the supernatant;
17. Add 2 ml PBS wash buffer to resuspend the cells, then 1000 rpm centrifugation for 5 minutes, discard the supernatant;
18. Add 0.5 ml PBS wash buffer to resuspend the cells and detect by flow cytometry (sample should be determined on the day on the machine and can also be added fixation overnight at 4�C then measured).
[PBS wash buffer: PBS +1% FBS +0.1% NaN3; Cat.:E16- FXP005]
[Cell fixation: 2% formaldehyde solution]
Notices: 1. This reagent has been pre-diluted for use at the recommended Volume per Test;
2. We typically use 1 X 106 cells in a 100 μl experimental sample (a test);
3. Since applications vary, each investigator should titrate the reagent to obtain optimal results;
4. 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;
5. If the sample can not be timely analyzed, please fix;
6. For research use only.