anti-Human IL-17A Monoclonal Antibody | anti-IL-17A antibody
Mouse anti-Human IL-17A Monoclonal Antibody, Purified
Gene Names
IL17A; IL17; CTLA8; IL-17; CTLA-8; IL-17A
Reactivity
Human
Applications
Flow Cytometry
Synonyms
IL-17A; Monoclonal Antibody; Mouse anti-Human IL-17A Monoclonal Antibody; Purified; anti-IL-17A antibody
Reactivity
Human
Clonality
Monoclonal
Isotype
Mouse IgG1, κ
Clone Number
64CAP17
Form/Format
Phosphate-buffered solution, pH 7.4, containing 0.09% sodium azide.
Concentration
0.5 mg/ml (varies by lot)
Sequence Length
155
Applicable Applications for anti-IL-17A antibody
ICFC
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]
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
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
Preparation and Storage
Store at 4°C. DO NOT FREEZE. LIGHT SENSITIVE MATERIAL.
Related Product Information for anti-IL-17A antibody
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.
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
NCBI Official Full Name
interleukin-17A
NCBI Official Synonym Full Names
interleukin 17A
NCBI Official Symbol
IL17A
NCBI Official Synonym Symbols
IL17; CTLA8; IL-17; CTLA-8; IL-17A
NCBI Protein Information
interleukin-17A
UniProt Protein Name
Interleukin-17A
UniProt Gene Name
IL17A
UniProt Synonym Gene Names
CTLA8; IL17; IL-17; IL-17A; CTLA-8
NCBI Description
The protein encoded by this gene is a proinflammatory cytokine produced by activated T cells. This cytokine regulates the activities of NF-kappaB and mitogen-activated protein kinases. This cytokine can stimulate the expression of IL6 and cyclooxygenase-2 (PTGS2/COX-2), as well as enhance the production of nitric oxide (NO). High levels of this cytokine are associated with several chronic inflammatory diseases including rheumatoid arthritis, psoriasis and multiple sclerosis. [provided by RefSeq, Jul 2008]
Uniprot Description
Ligand for IL17RA and IL17RC (PubMed:17911633). The heterodimer formed by IL17A and IL17F is a ligand for the heterodimeric complex formed by IL17RA and IL17RC (PubMed:18684971). Involved in inducing stromal cells to produce proinflammatory and hematopoietic cytokines (PubMed:8676080).
Research Articles on IL-17A
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Product Notes
The IL-17A il17a (Catalog #AAA8505777) 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. AAA Biotech'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. It is sometimes possible for the material contained within the vial of "IL-17A, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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