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Product Info Summary
| SKU: | PB10016 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | IHC, WB, ELISA (Cap) |
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Product info
Product Name
Anti-Mast Cell Tryptase/TPSAB1 Antibody Picoband®
SKU/Catalog Number
PB10016
PB1058 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Mast Cell Tryptase/TPSAB1 Antibody Picoband® catalog # PB10016. Tested in ELISA, IHC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Mast Cell Tryptase/TPSAB1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB10016)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human Mast Cell Tryptase recombinant protein (Position: H65-P275). Human Mast Cell Tryptase shares 77% and 76.1% amino acid (aa) sequence identity with mouse and rat Mast Cell Tryptase, respectively.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB10016 is reactive to TPSAB1 in Human
Observed Molecular Weight
30 kDa
Calculated molecular weight
30.5 kDa
Background of TPSAB1
Tryptase alpha-1 and tryptase beta-1 are enzymes that in humans are encoded by the same TPSAB1 gene. 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. In addition, 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.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
PB10016 is guaranteed for IHC, WB, ELISA (Cap) Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
ELISA(Cap) , 1-5μg/ml, Human, -
Positive Control
WB: 293T whole cell
IHC: human lung cancer tissue, human breast cancer tissue, human intestinal cancer tissue
Validation Images & Assay Conditions
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Western blot analysis of Mast Cell Tryptase using anti-Mast Cell Tryptase antibody (PB10016).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: 293T whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Mast Cell Tryptase antigen affinity purified polyclonal antibody (Catalog # PB10016) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Mast Cell Tryptase at approximately 30 kDa. The expected band size for Mast Cell Tryptase is at 31 kDa.
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IHC analysis of Mast Cell Tryptase using anti-Mast Cell Tryptase antibody (PB10016).
Mast Cell Tryptase was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Mast Cell Tryptase Antibody (PB10016) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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IHC analysis of Mast Cell Tryptase using anti-Mast Cell Tryptase antibody (PB10016).
Mast Cell Tryptase was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Mast Cell Tryptase Antibody (PB10016) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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IHC analysis of Mast Cell Tryptase using anti-Mast Cell Tryptase antibody (PB10016).
Mast Cell Tryptase was detected in a paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Mast Cell Tryptase Antibody (PB10016) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Sandwich ELISA - Recombinant human Tryptase/TPSAB1,B2 protein standard curve.
Use in combination with reagents from Human Tryptase/TPSAB1,B2 ELISA Kit EZ-Set (DIY Antibody Pairs) (EZ0898).
Specific Publications For Anti-Mast Cell Tryptase/TPSAB1 Antibody Picoband® (PB10016)
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2 Customer Q&As for Anti-Mast Cell Tryptase/TPSAB1 Antibody Picoband®
Question
The customer want to know how the ELISA application is validated, including the strategy of ELISA method, if there was a another antibody used with the PB100016 as pairs, as well as the concentration and tested samples. Would you please help me with this?
Verified customer
Asked: 2020-09-28
Answer
Our lab performed indirect ELISA with PB10016. It could not be used as capture antibody and be paired with other antibodies. Please see the following protocol. 1. Diluent the recombinant protein solution with coating buffer (Boster Cat# AR1106-4) to the concentrations of 10ng/ml, 5ng/ml and 2.5ng/ml respectively. (Note: Concentrations of the recombinant protein solution should be determined by end users.) 2. Immediately coat a 96-well microplate (Boster Cat# AR1100) with 100uL of diluted recombinant protein solutions and incubate overnight at 4°C. 3. Wash plate 3 times with wash buffer (Boster Cat# AR0030/AR0031). 4. Block the plate with 150uL blocking buffer (Boster Cat# AR1106-1) into each well and incubate at room temperature for 2 hours. 5. Wash plate 3 times with wash buffer (Boster Cat# AR0030/AR0031). 6. Diluent the primary antibody with antibody diluent buffer (Boster Cat# AR1106-2) to the working concentration of 0.1-0.5ug/ml. Add 100ul primary antibody into each well and incubate at 37°C for 1 hour. 7. Wash plate 3 times with wash buffer (Boster Cat# AR0030/AR0031). 8. Add 100μL of HRP conjugated goat anti-rabbit secondary antibody (Boster Cat# BA1054), diluted in diluent buffer (Boster Cat# AR1106-3) in a ratio of 1:5000, to each well. And incubate at 37°C for 30 minutes. 9. Wash plate 5 times with wash buffer (Boster Cat# AR0030/AR0031). 10. Add prepared TMB substrate solution(Boster Cat# AR1104) and incubate at 37℃ in dark for 30 minutes. 11. Add prepared TMB stop solution (Boster Cat# AR1105). 12. Read the O.D. absorbance at 450nm in a microplate reader within 30 min after adding the stop solution.
Boster Scientific Support
Answered: 2020-09-29
Question
We are currently using anti-Mast Cell Tryptase/TPSAB1 antibody PB10016 for human tissue, and we are happy with the IHC results. The species of reactivity given in the datasheet says human. Is it true that the antibody can work on primate tissues as well?
Verified Customer
Verified customer
Asked: 2017-11-20
Answer
The anti-Mast Cell Tryptase/TPSAB1 antibody (PB10016) has not been tested for cross reactivity specifically with primate tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in primate you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-11-20
