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Human IL-36A / IL-36 alpha ELISA Kit

SKU:
HUFI01718
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9UHA7
Sensitivity:
0.094ng/ml
Range:
0.156-10ng/ml
ELISA Type:
Sandwich
Synonyms:
IL36A, Interleukin-36 alpha, FIL1 epsilon, IL-1E, IL1F6, FIL1, FIL1 epsilon, FIL1, EPSILON, FIL1E, IL-1 epsilon, IL1, EPSILON, IL1E, IL1F6, IL-1F6, IL-1F6, FIL-1-epsilon, interleukin 1 family, member 6, epsilon, interleukin 1, epsilon, Interleukin 36
Reactivity:
Human
Research Area:
Immunology
Signaling Molecule:
Cytokine
€599
Frequently bought together:

Description

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Human IL-36A/IL-36 alpha ELISA Kit

The Human IL36A (IL36 Alpha) ELISA Kit is a highly reliable and sensitive assay designed for the quantitative detection of IL36 Alpha levels in human serum, plasma, and cell culture supernatants. With its high specificity, this kit provides accurate and reproducible results, making it an excellent choice for a variety of research applications.IL36 Alpha, also known as Interleukin-36 Alpha, is a pro-inflammatory cytokine that plays a crucial role in the immune response and inflammation regulation. Dysregulation of IL36 Alpha has been implicated in various inflammatory diseases, such as psoriasis and inflammatory bowel disease, making it an essential biomarker for studying and understanding these conditions.

The Human IL36A ELISA Kit from Assay Genie is a valuable tool for researchers looking to investigate the role of IL36 Alpha in disease pathogenesis and potential therapeutic targets. Its ease of use, high sensitivity, and specificity make it a valuable resource for studying the complex functions of IL36 Alpha in various research settings.

Product Name:Human IL-36A / IL-36 alpha ELISA Kit
Product Code:HUFI01718
Size:96 Assays
Alias:IL36A, Interleukin-36 alpha, FIL1 epsilon, IL-1E, IL1F6, FIL1, FIL1 epsilon, FIL1, EPSILON, FIL1E, IL-1 epsilon, IL1, EPSILON, IL1E, IL1F6, IL-1F6, IL-1F6, FIL-1-epsilon, interleukin 1 family, member 6, epsilon, interleukin 1, epsilon, Interleukin 36, Alpha, Interleukin-1 epsilon, interleukin-1 family member 6, Interleukin-36 Alpha
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human IL36A concentrations in serum plasma and other biological fluids.
Sensitivity:0.094ng/ml
Range:0.156-10ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human IL36A and the recovery rates were calculated by comparing the measured value to the expected amount of Human IL36A in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)86-10498
EDTA plasma(n=5)86-10192
UFH plasma(n=5)95-10297
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human IL36A 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%92-104%87-103%
EDTA plasma(n=5)83-98%83-96%82-99%
UFH plasma(n=5)85-98%80-100%81-89%
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
UniprotQ9UHA7
UniProt Protein Function:IL1F6: Belongs to the IL-1 family.
UniProt Protein Details:Protein type:Cytokine

Chromosomal Location of Human Ortholog: 2q12-q14.1

Cellular Component: extracellular region; extracellular space

Molecular Function:cytokine activity; interleukin-1 receptor binding

Biological Process: cytokine and chemokine mediated signaling pathway

NCBI Summary:The protein encoded by this gene is a cytokine that can activate NF-kappa-B and MAPK signaling pathways to generate an inflammatory response. The encoded protein functions primarily in skin and demonstrates increased expression in psoriasis. In addition, decreased expression of this gene has been linked to a poor prognosis in both hepatocellular carcinoma and colorectal cancer patients. [provided by RefSeq, Nov 2015]
UniProt Code:Q9UHA7
NCBI GenInfo Identifier:25008601
NCBI Gene ID:27179
NCBI Accession:Q9UHA7.1
UniProt Secondary Accession:Q9UHA7,Q53SR7, Q5BLR4, Q7RTZ8, B2RAD9,
UniProt Related Accession:Q9UHA7
Molecular Weight:17,684 Da
NCBI Full Name:Interleukin-36 alpha
NCBI Synonym Full Names:interleukin 36, alpha
NCBI Official Symbol:IL36A
NCBI Official Synonym Symbols:FIL1; FIL1E; IL1F6; IL-1F6; IL1(EPSILON); FIL1(EPSILON)
NCBI Protein Information:interleukin-36 alpha
UniProt Protein Name:Interleukin-36 alpha
UniProt Synonym Protein Names:FIL1 epsilon; Interleukin-1 epsilon; IL-1 epsilon; Interleukin-1 family member 6; IL-1F6
Protein Family:Interleukin
UniProt Gene Name:IL36A
UniProt Entry Name:IL36A_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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