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Human IL-15RA / IL-15 R alpha ELISA Kit

SKU:
HUFI00903
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q13261
Sensitivity:
4.688pg/ml
Range:
7.813-500pg/ml
ELISA Type:
Sandwich
Synonyms:
IL15RA, CD215, Interleukin-15 receptor subunit alpha, IL-15 R alpha, IL-15 receptor subunit alpha, IL-15RA, IL-15R-alpha, interleukin 15 receptor, alpha
Reactivity:
Human
Research Area:
Cell Biology
€599
Frequently bought together:

Description

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Human IL-15RA/IL-15 R alpha ELISA Kit

The Human IL15RA (IL-15R Alpha) ELISA Kit is a reliable and accurate tool for detecting levels of IL-15R Alpha in human samples such as serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reproducible results for a variety of research applications.IL-15R Alpha is a critical receptor involved in the immune response, specifically in the activation and proliferation of natural killer cells and CD8+ T cells. Dysregulation of IL-15R Alpha signaling has been linked to various autoimmune diseases, inflammatory disorders, and infectious diseases, making it a valuable biomarker for studying these conditions and potential therapeutic interventions.

By utilizing the Human IL15RA ELISA Kit, researchers can gain valuable insights into the role of IL-15R Alpha in immune function and disease pathogenesis, advancing our understanding of immune-mediated disorders and facilitating the development of targeted therapies.

Product Name:Human IL-15RA / IL-15 R alpha ELISA Kit
Product Code:HUFI00903
Size:96 Assays
Alias:IL15RA, CD215, Interleukin-15 receptor subunit alpha, IL-15 R alpha, IL-15 receptor subunit alpha, IL-15RA, IL-15R-alpha, interleukin 15 receptor, alpha
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human IL15RA concentrations in serum plasma and other biological fluids.
Sensitivity:4.688pg/ml
Range:7.813-500pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human IL15RA and the recovery rates were calculated by comparing the measured value to the expected amount of Human IL15RA in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)85-10395
EDTA plasma(n=5)85-10596
UFH plasma(n=5)91-10598
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human IL15RA and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)90-105%86-103%85-102%
EDTA plasma(n=5)85-96%85-101%87-101%
UFH plasma(n=5)81-88%80-95%80-98%
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
UniprotQ13261
UniProt Protein Function:IL15RA: High-affinity receptor for interleukin-15. Can signal both in cis and trans where IL15R from one subset of cells presents IL15 to neighboring IL2RG-expressing cells. Expression of different isoforms may alter or interfere with signal transduction. Isoform 5, isoform 6, isoform 7 and isoform 8 do not bind IL15. Signal transduction involves STAT3, STAT5, STAT6, JAK2 and SYK. 8 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:Receptor, cytokine; Membrane protein, integral

Chromosomal Location of Human Ortholog: 10p15.1

Cellular Component: plasma membrane

Molecular Function:hematopoietin/interferon-class (D200-domain) cytokine receptor activity; protein binding; signal transducer activity

Biological Process: cell proliferation; signal transduction

NCBI Summary:This gene encodes a cytokine receptor that specifically binds interleukin 15 (IL15) with high affinity. The receptors of IL15 and IL2 share two subunits, IL2R beta and IL2R gamma. This forms the basis of many overlapping biological activities of IL15 and IL2. The protein encoded by this gene is structurally related to IL2R alpha, an additional IL2-specific alpha subunit necessary for high affinity IL2 binding. Unlike IL2RA, IL15RA is capable of binding IL15 with high affinity independent of other subunits, which suggests distinct roles between IL15 and IL2. This receptor is reported to enhance cell proliferation and expression of apoptosis inhibitor BCL2L1/BCL2-XL and BCL2. Multiple alternatively spliced transcript variants of this gene have been reported.[provided by RefSeq, Apr 2010]
UniProt Code:Q13261
NCBI GenInfo Identifier:59799763
NCBI Gene ID:3601
NCBI Accession:Q13261.1
UniProt Secondary Accession:Q13261,Q3B769, Q5JVA1, Q5JVA2, Q5JVA4, Q6B0J2, Q7LDR4 Q7Z609, B4E2C2,
UniProt Related Accession:Q13261
Molecular Weight:24,388 Da
NCBI Full Name:Interleukin-15 receptor subunit alpha
NCBI Synonym Full Names:interleukin 15 receptor subunit alpha
NCBI Official Symbol:IL15RA
NCBI Official Synonym Symbols:CD215
NCBI Protein Information:interleukin-15 receptor subunit alpha
UniProt Protein Name:Interleukin-15 receptor subunit alpha
UniProt Synonym Protein Names:CD_antigen: CD215
Protein Family:Interleukin-15 receptor
UniProt Gene Name:IL15RA
UniProt Entry Name:I15RA_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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