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Human IL-27 / EBI3 ELISA Kit

SKU:
HUFI02094
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q14213
Sensitivity:
18.75pg/ml
Range:
31.25-2000pg/ml
ELISA Type:
Sandwich
Synonyms:
EBI3, Epstein-Barr virus-induced gene 3 protein, EBV-induced gene 3 protein, IL-27 subunit beta, IL-27B, IL-27, IL-27 p28 subunit,
Reactivity:
Human
Signaling Molecule:
Cytokine
$719
Frequently bought together:

Description

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Human IL-27/EBI3 ELISA Kit

The Human IL27 (Ebi3) ELISA Kit is specifically designed for the precise and reliable measurement of IL27 levels in human samples including serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring accurate and reproducible results for a variety of research applications.IL27, also known as Ebi3, is a key cytokine that plays a critical role in regulating immune responses and inflammation. It has been implicated in various diseases such as autoimmune disorders, infectious diseases, and cancer, making it a valuable biomarker for studying immune system function and disease progression.

With its advanced technology and user-friendly protocol, the Human IL27 (Ebi3) ELISA Kit provides researchers with a powerful tool for investigating the role of IL27 in health and disease, ultimately leading to new insights and potential therapeutic strategies.

Product Name:Human IL-27 / EBI3 ELISA Kit
Product Code:HUFI02094
Size:96 Assays
Alias:EBI3, Epstein-Barr virus-induced gene 3 protein, EBV-induced gene 3 protein, IL-27 subunit beta, IL-27B, IL-27, IL-27 p28 subunit
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human EBI3 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 EBI3 and the recovery rates were calculated by comparing the measured value to the expected amount of Human EBI3 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)85-10494
EDTA plasma(n=5)86-10295
UFH plasma(n=5)86-10095
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human EBI3 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-105%85-102%90-102%
EDTA plasma(n=5)87-96%83-101%83-98%
UFH plasma(n=5)81-92%81-98%82-100%
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
UniprotQ14213
UniProt Protein Function:IL27-beta: Cytokine with pro- and anti-inflammatory properties, that can regulate T-helper cell development, suppress T-cell proliferation, stimulate cytotoxic T-cell activity, induce isotype switching in B-cells, and that has diverse effects on innate immune cells. Among its target cells are CD4 T-helper cells which can differentiate in type 1 effector cells (TH1), type 2 effector cells (TH2) and IL17 producing helper T-cells (TH17). It drives rapid clonal expansion of naive but not memory CD4 T-cells. It also strongly synergizes with IL-12 to trigger interferon- gamma/IFN-gamma production of naive CD4 T-cells, binds to the cytokine receptor WSX-1/TCCR. Another important role of IL27 is its antitumor activity as well as its antiangiogenic activity with activation of production of antiangiogenic chemokines. Belongs to the type I cytokine receptor family. Type 3 subfamily.
UniProt Protein Details:Protein type:Secreted, signal peptide; Secreted

Chromosomal Location of Human Ortholog: 19p13.3

Cellular Component: extracellular region; extracellular space; plasma membrane

Molecular Function:cytokine activity; hematopoietin/interferon-class (D200-domain) cytokine receptor activity; interleukin-27 receptor binding; protein binding

Biological Process: cytokine and chemokine mediated signaling pathway; humoral immune response; positive regulation of alpha-beta T cell proliferation; positive regulation of interferon-gamma biosynthetic process; T cell proliferation; T-helper 1 type immune response

NCBI Summary:This gene was identified by its induced expression in B lymphocytes in response Epstein-Barr virus infection. It encodes a secreted glycoprotein belonging to the hematopoietin receptor family, and heterodimerizes with a 28 kDa protein to form interleukin 27 (IL-27). IL-27 regulates T cell and inflammatory responses, in part by activating the Jak/STAT pathway of CD4+ T cells. [provided by RefSeq, Sep 2008]
UniProt Code:Q14213
NCBI GenInfo Identifier:47605806
NCBI Gene ID:10148
NCBI Accession:Q14213.2
UniProt Secondary Accession:Q14213,O75269, A0N0N2,
UniProt Related Accession:Q14213
Molecular Weight:25,396 Da
NCBI Full Name:Interleukin-27 subunit beta
NCBI Synonym Full Names:Epstein-Barr virus induced 3
NCBI Official Symbol:EBI3 Â 
NCBI Official Synonym Symbols:IL27B; IL-27B Â 
NCBI Protein Information:interleukin-27 subunit beta
UniProt Protein Name:Interleukin-27 subunit beta
UniProt Synonym Protein Names:Epstein-Barr virus-induced gene 3 protein; EBV-induced gene 3 protein
Protein Family:Interleukin
UniProt Gene Name:EBI3 Â 
UniProt Entry Name:IL27B_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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