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Human Neurokinin B / NKB ELISA Kit

SKU:
HUFI02664
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9UHF0
Sensitivity:
9.375pg/ml
Range:
15.625-1000pg/ml
ELISA Type:
Competitive
Synonyms:
NKB
Reactivity:
Human
€599
Frequently bought together:

Description

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Human Neurokinin B/NKB ELISA Kit

The Human Neurokinin B (NKB) ELISA Kit is a highly sensitive and specific assay for the accurate quantification of NKB levels in human samples such as serum, plasma, and cell culture supernatants. This kit provides reliable and reproducible results, making it suitable for a variety of research applications.Neurokinin B is a neuropeptide that plays a crucial role in various physiological processes, such as regulating neurotransmitter release and modulating pain perception. Imbalances in NKB levels have been associated with conditions such as chronic pain, neurological disorders, and reproductive dysfunction, making it a valuable biomarker for studying these conditions and evaluating potential therapeutic interventions.

The Human NKB ELISA Kit from Assaygenie offers researchers a powerful tool for studying the role of NKB in health and disease, providing accurate and precise measurements to further the understanding of NKB-mediated pathways and potential treatment strategies.

Product Name:Human Neurokinin B / NKB ELISA Kit
Product Code:HUFI02664
Size:96 Assays
Alias:NKB
Detection method:Competitive ELISA, Coated with Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human NKB concentrations in serum plasma and other biological fluids.
Sensitivity:9.375pg/ml
Range:15.625-1000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human NKB and the recovery rates were calculated by comparing the measured value to the expected amount of Human NKB in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)88-10496
EDTA plasma(n=5)85-10191
UFH plasma(n=5)88-9894
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human NKB and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)85-104%89-105%95-104%
EDTA plasma(n=5)87-95%82-101%82-90%
UFH plasma(n=5)80-95%82-99%82-92%
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)60ul4°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 pipettetips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotQ9UHF0
UniProt Protein Function:TAC3: Tachykinins are active peptides which excite neurons, evoke behavioral responses, are potent vasodilators and secretagogues, and contract (directly or indirectly) many smooth muscles. Belongs to the tachykinin family. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:Secreted; Secreted, signal peptide

Chromosomal Location of Human Ortholog: 12q13.3

Cellular Component: extracellular region; extracellular space

Molecular Function:receptor binding

Biological Process: female pregnancy

Disease: Hypogonadotropic Hypogonadism 10 With Or Without Anosmia

NCBI Summary:This gene encodes a member of the tachykinin family of secreted neuropeptides. The encoded preproprotein is proteolytically processed to generate the mature peptide, which is primarily expressed in the central and peripheral nervous systems and functions as a neurotransmitter. This peptide is the ligand for the neurokinin-3 receptor. This protein is also expressed in the outer syncytiotrophoblast of the placenta and may be associated with pregnancy-induced hypertension and pre-eclampsia. Mutations in this gene are associated with normosmic hypogonadotropic hypogonadism. Alternative splicing results in multiple transcript variants, at least one of which encodes an isoform that is proteolytically processed. [provided by RefSeq, Feb 2016]
UniProt Code:Q9UHF0
NCBI GenInfo Identifier:18203501
NCBI Gene ID:6866
NCBI Accession:Q9UHF0.1
UniProt Secondary Accession:Q9UHF0,Q6IAG2, Q71BC6, Q71BC9,
UniProt Related Accession:Q9UHF0
Molecular Weight:11,358 Da
NCBI Full Name:Tachykinin-3
NCBI Synonym Full Names:tachykinin 3
NCBI Official Symbol:TAC3  
NCBI Official Synonym Symbols:NKB; HH10; NKNB; PRO1155; ZNEUROK1  
NCBI Protein Information:tachykinin-3
UniProt Protein Name:Tachykinin-3
UniProt Synonym Protein Names:ZNEUROK1
Protein Family:Tachykinin
UniProt Gene Name:TAC3  
UniProt Entry Name:TKNK_HUMAN

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

Equilibrate the TMB substrate for at least 30 min at 37°C beforeuse. When diluting samples and reagents, they must be mixed completely andevenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coatedplate respectively, and then, record their positions. It isrecommended to measure each standard and sample in duplicate. Washplate 2 times before adding standard, sample and control (zero) wells!
2.Add Sample and Biotin-detection antibody: Add 50µL of Standard, Blank or Sample per well. The blankwell is added with Sample Dilution Buffer. Immediately add 50 µL of biotin-labelled antibody workingsolution to each well. Cover with the plate sealer provided. Gently tap the plate to ensure thoroughmixing. Incubate for 45 minutes at 37°C. (Solutions are added to the bottom of micro-ELISA platewell, avoid touching plate walls and foaming).
3.Wash: Aspirate each well and wash, repeating the process three timesWash by filling each well with Wash Buffer (approximately 350µL)using a squirt bottle, multi-channel pipette, manifold dispenser orautomated washer. Complete removal of liquid at each step is essentialto good performance. After the last wash, remove any remaining WashBuffer by aspirating or decanting. Invert the plate and pat it againstthick clean absorbent paper.
4.HRP-Streptavidin Conjugate(SABC): Add 100µL of SABC workingsolution to each well. Cover with a new Plate sealer. Incubate for30minutes at 37°C.
5.Wash: Repeat the aspiration/wash process for five times.
6.TMB Substrate: Add 90µL of TMB Substrate to each well. Coverwith a new Plate sealer. Incubate for about 10-20 minutes at 37°C.Protect from light. The reaction time can be shortened or extendedaccording to the actual color change, but not more than 30minutes.When apparent gradient appeared in standard wells, you can terminatethe reaction.
7.Stop: Add 50µL of Stop Solution to each well. Color turn toyellow immediately. The adding order of stop solution should be as thesame as the substrate solution.
8.OD Measurement: Determine the optical density (OD Value) of each wellat once, using a microplate reader set to 450 nm. You should open themicroplate reader ahead, preheat the instrument, and set the testing parameters.

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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