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Product Info Summary
| SKU: | PB9539 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-COMT Antibody Picoband®
SKU/Catalog Number
PB9539
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-COMT Antibody Picoband® catalog # PB9539. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human, Mouse, Rat. 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-COMT Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9539)
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 COMT recombinant protein (Position: G52-P271). Human COMT shares 81.9% and 81% amino acid (aa) sequence identity with mouse and rat COMT, respectively.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9539 is reactive to COMT in Human, Mouse, Rat
Observed Molecular Weight
26 kDa
Calculated molecular weight
30.0 kDa
Background of COMT
Catechol O-methyltransferase, also called COMT, is one of the major mammalian enzymes involved in the metabolic degradation of catecholamines. This gene is mapped to 22q11.21. Catechol-O-methyltransferase catalyzes the transfer of a methyl group from S-adenosylmethionine to catecholamines, including the neurotransmitters dopamine, epinephrine, and norepinephrine. This O-methylation results in one of the major degradative pathways of the catecholamine transmitters. In addition to its role in the metabolism of endogenous substances, COMT is important in the metabolism of catechol drugs used in the treatment of hypertension, asthma, and Parkinson disease. COMT is found in two forms in tissues, a soluble form (S-COMT) and a membrane-bound form (MB-COMT). The differences between S-COMT and MB-COMT reside within the N-termini.
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
PB9539 is guaranteed for Flow Cytometry, IHC, WB 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, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human SH-SY5Y whole cell, human HepG2 whole cell, rat brain tissue, rat liver tissue, rat C6 whole cell, mouse brain tissue, mouse liver tissue, mouse Neuro-2a whole cell
IHC: Mhuman human breast cancer tissue
FCM: U251 Cell
Validation Images & Assay Conditions
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Western blot analysis of COMT using anti-COMT antibody (PB9539).
Electrophoresis was performed on a 13% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human SH-SY5Y whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: rat brain tissue lysates,
Lane 4: rat liver tissue lysates,
Lane 5: rat C6 whole cell lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse Neuro-2a 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-COMT antigen affinity purified polyclonal antibody (Catalog # PB9539) 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 ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for COMT at approximately 26 kDa. The expected band size for COMT is at 30 kDa.
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IHC analysis of COMT using anti-COMT antibody (PB9539).
COMT was detected in a paraffin-embedded section of human 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 2 μg/ml rabbit anti-COMT Antibody (PB9539) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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Flow Cytometry analysis of U251 cells using anti-COMT antibody (PB9539).
Overlay histogram showing U251 cells stained with PB9539 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-COMT Antibody (PB9539, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-COMT Antibody Picoband® (PB9539)
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1 Customer Q&As for Anti-COMT Antibody Picoband®
Question
We are currently using anti-COMT antibody PB9539 for rat tissue, and we are satisfied with the IHC results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on primate tissues as well?
B. Li
Verified customer
Asked: 2017-03-20
Answer
The anti-COMT antibody (PB9539) 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: 2017-03-20
