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Human FADS2 / Fatty acid desaturase 2 ELISA Kit

SKU:
HUFI00661
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
O95864
Sensitivity:
46.875pg/ml
Range:
78.125-5000pg/ml
ELISA Type:
Sandwich
Synonyms:
FADS2, Fatty acid desaturase 2, D6D, DES6, FADSD6, LLCDL2, SLL0262, TU13
Reactivity:
Human
Research Area:
Cell Biology
$719
Frequently bought together:

Description

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Human FADS2/Fatty acid desaturase 2 ELISA Kit

The Human FADS2 (Fatty Acid Desaturase 2) ELISA Kit is specifically designed for the quantitative measurement of FADS2 levels in human samples such as serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit provides accurate and reliable results, making it a valuable tool for various research applications. FADS2 is an important enzyme involved in the biosynthesis of long-chain polyunsaturated fatty acids, playing a key role in maintaining lipid homeostasis and cell membrane function.

Dysregulation of FADS2 has been linked to various diseases such as cardiovascular disorders, metabolic syndromes, and inflammatory conditions, highlighting its significance as a potential biomarker for disease detection and treatment development. Overall, the Human FADS2 ELISA Kit offers researchers a precise and efficient method for studying the role of FADS2 in human health and disease, paving the way for new insights and therapeutic interventions in the field of lipid metabolism and related disorders.

Product Name:Human FADS2 / Fatty acid desaturase 2 ELISA Kit
Product Code:HUFI00661
Size:96 Assays
Alias:FADS2, Fatty acid desaturase 2, D6D, DES6, FADSD6, LLCDL2, SLL0262, TU13
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human FADS2 concentrations in serum plasma and other biological fluids.
Sensitivity:46.875pg/ml
Range:78.125-5000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human FADS2 and the recovery rates were calculated by comparing the measured value to the expected amount of Human FADS2 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)89-10296
EDTA plasma(n=5)85-9893
UFH plasma(n=5)87-10094
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human FADS2 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)89-102%89-103%87-104%
EDTA plasma(n=5)87-99%83-97%86-97%
UFH plasma(n=5)81-94%81-100%87-97%
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
UniprotO95864
UniProt Protein Function:FADS2: Component of a lipid metabolic pathway that catalyzes biosynthesis of highly unsaturated fatty acids (HUFA) from precursor essential polyunsaturated fatty acids (PUFA) linoleic acid (LA) (18:2n-6) and alpha-linolenic acid (ALA) (18:3n-3). Catalyzes the first and rate limiting step in this pathway which is the desaturation of LA (18:2n-6) and ALA (18:3n-3) into gamma- linoleic acid (GLA) (18:3n-6) and stearidonic acid (18:4n-3) respectively and other desaturation steps. Highly unsaturated fatty acids (HUFA) play pivotal roles in many biological functions. It catalizes as well the introduction of a cis double bond in palmitate to produce the mono-unsaturated fatty acid sapienate, the most abundant fatty acid in sebum. Belongs to the fatty acid desaturase family. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:EC 1.14.19.-; Lipid Metabolism - alpha-linolenic acid; Lipid Metabolism - unsaturated fatty acid biosynthesis; Membrane protein, integral; Membrane protein, multi-pass; Oxidoreductase

Chromosomal Location of Human Ortholog: 11q12.2

Cellular Component: endoplasmic reticulum membrane; integral to plasma membrane; membrane

Molecular Function:linoleoyl-CoA desaturase activity

Biological Process: linoleic acid metabolic process

NCBI Summary:The protein encoded by this gene is a member of the fatty acid desaturase (FADS) gene family. Desaturase enzymes regulate unsaturation of fatty acids through the introduction of double bonds between defined carbons of the fatty acyl chain. FADS family members are considered fusion products composed of an N-terminal cytochrome b5-like domain and a C-terminal multiple membrane-spanning desaturase portion, both of which are characterized by conserved histidine motifs. This gene is clustered with family members at 11q12-q13.1; this cluster is thought to have arisen evolutionarily from gene duplication based on its similar exon/intron organization. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Jul 2013]
UniProt Code:O95864
NCBI GenInfo Identifier:74762131
NCBI Gene ID:9415
NCBI Accession:O95864.1
UniProt Secondary Accession:O95864,Q6MZQ7, Q96H07, Q96SV8, Q9H3G3, Q9Y3X4, A8K2M6 B7Z634,
UniProt Related Accession:O95864
Molecular Weight:52kDa
NCBI Full Name:Fatty acid desaturase 2
NCBI Synonym Full Names:fatty acid desaturase 2
NCBI Official Symbol:FADS2
NCBI Official Synonym Symbols:D6D; DES6; TU13; FADSD6; LLCDL2; SLL0262
NCBI Protein Information:fatty acid desaturase 2
UniProt Protein Name:Fatty acid desaturase 2
UniProt Synonym Protein Names:Acyl-CoA 6-desaturase; Delta(6) fatty acid desaturase; D6D; Delta(6) desaturase; Delta-6 desaturase
Protein Family:Fatty acid desaturase
UniProt Gene Name:FADS2

*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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