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Human IL-17 ELISA Kit

SKU:
HUFI02585
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q16552
Sensitivity:
18.75pg/ml
Range:
31.25-2000pg/ml
ELISA Type:
Sandwich
Synonyms:
IL-17, Interleukin 17, CTLA-8, IL17A, IL-17CTLA-8, IL17interleukin-17A, interleukin 17A
Reactivity:
Human
Research Area:
Cell Biology
Signaling Molecule:
Cytokine
€599
Frequently bought together:

Description

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Human IL-17 ELISA Kit

The Human IL-17 (Interleukin-17) ELISA Kit is specifically designed for the quantitative detection of IL-17 levels in human samples such as serum, plasma, and cell culture supernatants. This kit is known for its high sensitivity and specificity, providing accurate and reliable results for a variety of research applications.IL-17 is a key cytokine involved in the regulation of immune responses and inflammation. Elevated levels of IL-17 have been linked to various autoimmune diseases, including rheumatoid arthritis, psoriasis, and multiple sclerosis.

Understanding the level of IL-17 in biological samples can help researchers gain insight into the pathogenesis of these diseases and potentially lead to the development of targeted therapies.With its easy-to-use format and high-quality components, the Human IL-17 ELISA Kit from Assay Genie is a valuable tool for researchers studying immune responses, inflammation, and autoimmune diseases. Get reliable and reproducible results with this kit, making it a must-have for laboratories focusing on immunology and inflammatory research.

Product Name:Human IL-17 ELISA Kit
Product Code:HUFI02585
Size:96 Assays
Alias:IL-17, Interleukin 17, CTLA-8, IL17A, IL-17CTLA-8, IL17interleukin-17A, interleukin 17A
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human IL-17A concentrations in serum plasma and other biological fluids.
Sensitivity:18.75pg/ml
Range:31.25-2000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human IL-17A and the recovery rates were calculated by comparing the measured value to the expected amount of Human IL-17A in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)86-10495
EDTA plasma(n=5)91-10397
UFH plasma(n=5)86-10294
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human IL-17A and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)86-104%86-105%87-102%
EDTA plasma(n=5)88-96%88-96%87-101%
UFH plasma(n=5)84-97%93-99%81-100%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotQ16552
UniProt Protein Function:IL17A: Induces stromal cells to produce proinflammatory and hematopoietic cytokines. Enhances the surface expression of ICAM1/intracellular adhesion molecule 1 in fibroblasts. Belongs to the IL-17 family.
UniProt Protein Details:Protein type:Secreted; Secreted, signal peptide

Chromosomal Location of Human Ortholog: 6p12

Cellular Component: extracellular space; external side of plasma membrane

Molecular Function:cytokine activity

Biological Process: cell death; cell-cell signaling; apoptosis; positive regulation of interleukin-23 production; positive regulation of osteoclast differentiation; protein amino acid glycosylation; positive regulation of transcription from RNA polymerase II promoter; immune response; inflammatory response

NCBI Summary: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 Code:Q16552
NCBI GenInfo Identifier:2498481
NCBI Gene ID:3605
NCBI Accession:Q16552.1
UniProt Secondary Accession:Q16552,Q5T2P0,
UniProt Related Accession:Q16552
Molecular Weight:17,504 Da
NCBI Full Name:Interleukin-17A
NCBI Synonym Full Names:interleukin 17A
NCBI Official Symbol:IL17A  
NCBI Official Synonym Symbols:IL17; CTLA8; IL-17; IL-17A  
NCBI Protein Information:interleukin-17A; CTLA-8; cytotoxic T-lymphocyte-associated antigen 8; cytotoxic T-lymphocyte-associated protein 8; interleukin 17 (cytotoxic T-lymphocyte-associated serine esterase 8)
UniProt Protein Name:Interleukin-17A
UniProt Synonym Protein Names:Cytotoxic T-lymphocyte-associated antigen 8; CTLA-8
Protein Family:Interleukin
UniProt Gene Name:IL17A  
UniProt Entry Name:IL17_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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