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Human TPSAB1(Tryptase alpha/beta-1) ELISA Kit

SKU:
HUFI03369
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q15661
Sensitivity:
0.938ng/ml
Range:
1.563-100ng/ml
ELISA Type:
Sandwich
Synonyms:
Tryptase alpha, TPS1, TPSAB1, Tryptase 1, TPS2, TPSB1, tryptase alpha, beta 1
Reactivity:
Human
€599
Frequently bought together:

Description

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Human TPSAB1 (Tryptase alpha/beta-1) ELISA Kit

The Human TPSAB1 (Tryptase Alpha/Beta 1) ELISA Kit is a reliable and accurate tool for detecting TPSAB1 levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring consistent and reproducible results that are crucial for various research applications.TPSAB1, also known as Tryptase Alpha/Beta 1, is an important enzyme involved in immune responses and allergic reactions. It is a key marker for conditions such as asthma, allergies, and inflammatory disorders, making it a valuable biomarker for studying these diseases and potential therapeutic interventions.

By using the Human TPSAB1 ELISA Kit, researchers can gain valuable insights into the role of TPSAB1 in immune responses and allergic reactions, ultimately leading to a deeper understanding of related diseases and the development of targeted treatments.

Product Name:Human TPSAB1(Tryptase alpha/beta-1) ELISA Kit
Product Code:HUFI03369
Size:96 Assays
Alias:Tryptase alpha, TPS1, TPSAB1, Tryptase 1, TPS2, TPSB1, tryptase alpha, beta 1
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human TPSAB1 concentrations in serum plasma and other biological fluids.
Sensitivity:0.938ng/ml
Range:1.563-100ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human TPSAB1 and the recovery rates were calculated by comparing the measured value to the expected amount of Human TPSAB1 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)85-10293
EDTA plasma(n=5)89-9793
UFH plasma(n=5)85-10293
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human TPSAB1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)86-103%87-105%87-99%
EDTA plasma(n=5)84-99%84-94%82-97%
UFH plasma(n=5)95-100%80-94%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)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
UniprotQ15661
UniProt Protein Function:TPSAB1: Tryptase is the major neutral protease present in mast cells and is secreted upon the coupled activation-degranulation response of this cell type. Belongs to the peptidase S1 family. Tryptase subfamily. 2 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:Protein type:Secreted, signal peptide; EC 3.4.21.59; Protease; Secreted

Chromosomal Location of Human Ortholog: 16p13.3

Cellular Component: extracellular matrix; extracellular space; extracellular region

Molecular Function:protein binding; serine-type peptidase activity; serine-type endopeptidase activity

Biological Process: extracellular matrix disassembly; extracellular matrix organization and biogenesis; defense response; proteolysis

NCBI Summary:Tryptases comprise a family of trypsin-like serine proteases, the peptidase family S1. Tryptases are enzymatically active only as heparin-stabilized tetramers, and they are resistant to all known endogenous proteinase inhibitors. Several tryptase genes are clustered on chromosome 16p13.3. These genes are characterized by several distinct features. They have a highly conserved 3' UTR and contain tandem repeat sequences at the 5' flank and 3' UTR which are thought to play a role in regulation of the mRNA stability. These genes have an intron immediately upstream of the initiator Met codon, which separates the site of transcription initiation from protein coding sequence. This feature is characteristic of tryptases but is unusual in other genes. The alleles of this gene exhibit an unusual amount of sequence variation, such that the alleles were once thought to represent two separate genes, alpha and beta 1. Beta tryptases appear to be the main isoenzymes expressed in mast cells; whereas in basophils, alpha tryptases predominate. Tryptases have been implicated as mediators in the pathogenesis of asthma and other allergic and inflammatory disorders. [provided by RefSeq, Jul 2008]
UniProt Code:Q15661
NCBI GenInfo Identifier:18202508
NCBI Gene ID:7177
NCBI Accession:Q15661.1
UniProt Secondary Accession:Q15661,P15157, Q15663, Q6B052, Q9H2Y4, Q9H2Y5, Q9UQI1 D2E6R9, D2E6S1,
UniProt Related Accession:Q15661
Molecular Weight:Observed MW: 31kDaCalculated MW: 29kDa/30kDa
NCBI Full Name:Tryptase alpha/beta-1
NCBI Synonym Full Names:tryptase alpha/beta 1
NCBI Official Symbol:TPSAB1  
NCBI Official Synonym Symbols:TPS1; TPS2; TPSB1  
NCBI Protein Information:tryptase alpha/beta-1; tryptase I; tryptase-1; tryptase-I; tryptase-III; tryptase beta 1; tryptase beta I; tryptase beta-1; tryptase alpha-1; tryptase alpha II; mast cell beta I tryptase; mast cell alpha II tryptase
UniProt Protein Name:Tryptase alpha/beta-1
UniProt Synonym Protein Names:Tryptase I; Tryptase alpha-1
Protein Family:Tryptase
UniProt Gene Name:TPSAB1  
UniProt Entry Name:TRYB1_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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