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Human Tryptase alpha/beta-1 (TPSAB1) ELISA Kit (HUEB1052)

SKU:
HUEB1052
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q15661
Range:
0.156-10 ng/mL
ELISA Type:
Sandwich
Synonyms:
Tryptase alpha, TPS1, TPSAB1, Tryptase 1, TPS2, TPSB1, tryptase alpha, beta 1
Reactivity:
Human
€649
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Description

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

The Human Tryptase Alpha/Beta-1 (TPSAB1) ELISA Kit is specifically designed for the accurate and precise detection of TPSAB1 levels in human serum, plasma, and cell culture supernatants. This highly sensitive and specific kit guarantees reliable and reproducible results, making it an ideal tool for researchers in various fields.TPSAB1 is a protein involved in allergic and inflammatory responses, making it a key marker for conditions such as asthma, eczema, and allergic rhinitis. By measuring TPSAB1 levels, researchers can gain valuable insights into the underlying mechanisms of these diseases and potentially identify new therapeutic targets.

This ELISA kit offers a simple and efficient way to quantify TPSAB1 levels and study its role in various physiological and pathological conditions. With its high performance and user-friendly protocol, the Human Tryptase Alpha/Beta-1 ELISA Kit is a valuable tool for advancing research in the field of immunology and allergy.

Product Name:Human Tryptase alpha/beta-1 (TPSAB1) ELISA Kit
SKU:HUEB1052
Size:96T
Target:Human Tryptase alpha/beta-1 (TPSAB1)
Synonyms:Tryptase I, Tryptase alpha-1, Tryptase-1, TPS1, TPS2, TPSB1
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Human
Detection Range:0.156-10ng/mL
Sensitivity:0.06ng/mL
Intra CV:4.1%
Inter CV:7.9%
Linearity:
Sample1:21:41:81:16
Serum(N=5)97-107%110-120%109-120%106-116%
EDTA Plasma(N=5)90-100%97-105%102-112%96-108%
Heparin Plasma(N=5)112-120%115-125%86-98%100-109%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum10296-108
Plasma10498-110
Function:Tryptase is the major neutral protease present in mast cells and is secreted upon the coupled activation-degranulation response of this cell type. May play a role in innate immunity. Isoform 2 cleaves large substrates, such as fibronectin, more efficiently than isoform 1, but seems less efficient toward small substrates (PubMed:18854315).
Uniprot:Q15661
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant human Tryptase alpha/beta-1
Sub Unit:Homotetramer. The active tetramer is converted to inactive monomers at neutral and acidic pH in the absence of heparin. Low concentrations of inactive monomers become active monomers at pH 6.0 in the presence of heparin. When the concentration of active monomers is higher, they convert to active monomers and then to active tetramers. These monomers are active and functionally distinct from the tetrameric enzyme. In contrast to the hidden active sites in the tetrameric form, the active site of the monomeric form is accessible for macromolecular proteins and inhibitors eg: fibrinogen which is a substrate for the monomeric but not for the tetrameric form. The monomeric form forms a complex with SERPINB6.
Research Area:Immunology
Subcellular Location:Secreted Released from the secretory granules upon mast cell activation.
Storage:Please see kit components below for exact storage details
Note:For research use only
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
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

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

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

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

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 Type Protocol
Serum If 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.
Plasma Collect 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 Fluid Collect 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 supernatant Collect 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 lysates Solubilize 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 homogenates The 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 lysates Rinse 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 Milk Collect 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.
ELISA
Human Tryptase alpha / TPS1 ELISA Kit

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